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Data from: Methylation-based markers for the estimation of age in African Cheetah, Acinonyx jubatus
<p>This is a dataset for methylation analyses in cheetah done by EpiTYPER mass array.</p>
Supplementary File 7 from: Rapier-Sharman N et. al., Secondary Transcriptomic Analysis of Triple-Negative Breast Cancer Reveals Reliable Universal and Subtype-Specific Mechanistic Markers, 2024
<p>Supplementary Materials File 7. Please note that though the order of the supplementary materials has changed since initial upload (File S7 was previously File S9 or S10), the contents of this zipped folder remain the same.</p>
Fig. 1 in Problematic Biases in the Availability of Molecular Markers in Protists: The Example of the Dinoflagellates
Fig. 1. Number of species of the most speciose dinoflagellate genera (> 11 species per genus). The empty bars represented the number of described species based on Gómez (2012a). The black bars represent the number of species with, at least, one nucleotide sequence available in DDBJ/EMBL/GenBank in January 2013.
ASPH and Hypoxia Marker Expression in Head and Neck Carcinomas: Implications for HPV-Associated Tumours
<p><strong>Data open:</strong> file with parametres used for the multivariate evaluation. </p>
Metazoa-level USCOs as markers in species delimitation and classification
<p><span>Metazoa-level<strong> </strong>Universal Single-Copy Orthologs (USCOs) are universally applicable markers for DNA taxonomy in animals which can replace or supplement single-gene barcoding. While Metazoa-level USCOs from target enrichment data were shown to reliably distinguish species, it remains to be tested whether USCOs are an evenly distributed, representative sample of a given metazoan genome, and hence can facilitate detection of past hybridization events. Besides, unlinked loci are a principal assumption in coalescent-based species delimitation approaches. 239 chromosome-level genomes were analyzed to show that Metazoa-level<strong> </strong>USCOs are a representative sample of a genome: in terms of distances to each other on a chromosome, but also over the chromosomes, they are almost as evenly distributed as protein-coding genes in general are. We tested the suitability of Metazoa-level USCOs extracted from genomes for species delimitation and phylogeny in four case studies: <em>Anopheles</em> mosquitos, <em>Drosophila</em> fruit flies, <em>Heliconius </em>butterflies, and Darwin's finches. In almost all instances USCOs allowed delineating species and yielded phylogenies that correspond to those generated from whole genome data.<strong> </strong>Our results show<strong> </strong>that USCO genes can be considered as genetically unlinked for practical purposes and representative for an entire metazoan genome. Our phylogenetic analyses demonstrate that USCOs may complement single-gene barcoding and provide more accurate taxonomic inferences. Combining USCOs from sources that used different versions of ortholog reference libraries to infer marker orthology may be challenging and at times impact taxonomic conclusions. However, we expect this problem to become less severe as the size of genome reference libraries and their sampling of organismic lineages is rapidly increasing.</span></p>
The more the better: Fatty acids are predictive markers of honey bee, Apis mellifera, worker longevity
<p>Fatty acids (FA), stemming from nutrition, form triglycerides that are key components for insect energy reserves. In managed <em>Apis mellifera</em> colonies, supplementary feeding is common practice, yet micronutrients and microbiota (i.e. B-vitamins and probiotics) are often neglected. Given that B-vitamins are obligate cofactors for FA metabolism, and probiotics likely play key roles as well (i.e. <em>Lactobacillus</em> spp. synthesize B-vitamins), understanding how they contribute to FA acquisition remains unknown. Indeed, FAs are established predictors to <em>A. mellifera</em> longevity, and as such, are a logical point of interest in long-lived "winter" bees, where <em>A. mellifera</em> colony losses typically occur. Here, in a hoarding cage trial, freshly emerged adult winter workers were exposed to antibiotics (ABX) to decouple innate benefits associated to native gut microbiota, or left unexposed to ABX (N=72 cages, N=2088 experimental workers). Subsequently, all workers were fed different diets containing either probiotics, B-vitamins, with replicate treatments given <em>ad libitum</em> access to pollen (mimicking real-hive scenarios) or left blank (control). At the end of the trial, a subsample (n=356) had their total FA contents analyzed using <u>G</u>as <u>C</u>hromatography coupled to <u>F</u>lame <u>I</u>onization <u>D</u>etector (GC-FID). Irrespective of dietary treatment, every worker contained all 11 identified FAs, aligning our results with <em>a priori</em> evidence and highlighting their underlying key roles for bee physiology and health. We show for the first time that B-vitamins alone did not improve the overall abundance of individual FAs (g), yet significant differences were associated with presence/absence of bacteria and/or access to pollen, reconfirming likely ties of microbiota aiding in nutrient breakdown of complex polysaccharides found in pollen. Finally, of clear importance, there was a positive significant correlation between total lipid content and worker longevity (+2.4 median day lifespan increase / mg of FA), thereby confirming the relevance of FAs for honey bee worker longevity.</p>
Images of mouse VTA and SNc sections labeled for neurotransmitter phenotype markers via RNAscope
<p>48 coronal sections from 3 male and 3 female C57Bl/6J mice were labeled for mRNAs encoding the canonical vesicular transporters for dopamine (VMAT2), GABA (VGAT), and glutamate (VGLUT2). These sections were then imaged via confocal microscopy and saved as CZI files, editable with Zeiss Zen software. These image files include 'event marker' graphics, indicating where neurons positive for one or more of the above transporters are. </p> <p>The protocols used for sample preparation, labeling, imaging, and counting are linked in the Related Works section. </p> <p>Cell count data are organized by subregion (SUBREGION counts for R.csv), anterior-posterior (e.g. VGATVTA.csv), or anterior-posterior and medial-lateral distribution (e.g. 'snc triple.csv' or 'snc vmat2 AP and ML.csv').</p> <p>The R code provided uses these csv files to visualize expression patterns across VTA and SNc. </p>
Ditrect STORM imaging and image reconstruction of the transcription initiation marker P-S5 and nuclear PI(4,5)P2 indirectly immunolabeled with AF647 (red) and AF555 (green) in control cells.
<p>U-2 OS cells were grown in DMEM with 10% FBS at 37°C and 5% CO<sub>2</sub>. Cells were plated one day before staining in ~50% confluence on the high-precision 12 mm round coverslips treated with Hellmanex, sonicated, washed, dried and sterilized. Cells were control treated with 1:000 DMSO in the culture media.</p> <p>U2OS cells were washed twice with PBS (pH 7.4) and fixed for 30 min in 2% PFA in PBS, washed 3-times for 5 min with PBS, then permeabilized in 0.1% Triton X-100 in PBS for 20 min, washed 3-times for 5 min by PBS and blocked in filtered 5% BSA in PBS for 30 min. Cells were incubated for 45 min with rabbit polyclonal IgG anti-RNAPII CTD P-S5 (Abcam ab5131) 3 µg/mL and mouse ascites IgM anti-PI(4,5)P2 2C11 (Z-A045; Echelon Biosci. Inc., USA) 5 µg/mL in 5% BSA in PBS, washed 3-times for 5 min in PBS and incubated for 30 min with goat anti-mouse IgM (µ-chain) AF555 (Jackson ImmunoRes. A24126) 10 µg/mL; goat anti-rabbit IgG AF647 (Invitrogen A21245) 10 µg/mL diluted in 5% BSA in PBS. Then the cells were washed 3-times for 5 min in PBS, post-fixed for 15 min in 2% PFA in PBS and washed 3-times for 5 min in PBS. All procedures were performed at RT and the cells were stored in PBS in the fridge overnight prior imaging.</p> <p>Coverslips with cells were mounted in the Chamlide chamber (Live Cell Instrument, Korea) and covered with imaging buffer (PBS pH 7.4, 50 mM MEA). Single-molecule localizations (SMLs) data were acquired by Zeiss Elyra PS.1 equipped with HR Diode 642-150 and HR DPSS 561-200 lasers, Alpha Plan-Apochromat 100x/1.46 oil DIC M27 Elyra objective and Andor EM CCD iXon DU 897 camera and Zeiss ZEN Black 2.1 SP3 software (Zeiss). AF647 and AF555 photo-switching was achieved by HiLo illumination and TIRF HP FOV with 100% power of 642nm or 561nm laser, and the signal was acquired via MBS 642 + EF LP 655 and MBS 561 + EF BP 570-620 / LP 750 filters, respectively. Exposure time was 40 ms and EM gain was 300 for both channels.</p> <p>SMLs were calculated in 2D by Zeiss ZEN Black 2.1 SP3 software using x,y 2D Gauss fit with point spread function (PSF) half width 177.9 nm, peak mask size 9 pixels and peak intensity to noise 6 and accounted for overlap in 2D with max cluster size 10. SMLs were rendered in ZEN software with 10 nm/px resolution and 1x PSF expansion factor. The data were model-based drift corrected in ZEN. Two channels were aligned using tetraspec beads fiducial markers for affine calibration. Drift-corrected and aligned localization coordinates were exported as text files. Text files were converted into csv files and imported using self-written macro (Hoboth et al., 2021a) into the ImageJ2 (Rueden et al., 2017) plug-in ThunderSTORM, visualized by normalized Gaussian method (Ovesny et al., 2014).</p>
Ditrect STORM imaging and image reconstruction of the transcription elongation marker P-S2 and nuclear PI(3,4)P2 indirectly immunolabeled with AF647 (red) and AF555 (green) in THZ1 treated cells.
<p>U-2 OS cells were grown in DMEM with 10% FBS at 37°C and 5% CO<sub>2</sub>. Cells were plated one day before staining in ~50% confluence on the high-precision 12 mm round coverslips treated with Hellmanex, sonicated, washed, dried and sterilized. Cells were treated for 3h with 1 µM THZ1 (MedChem HY80013) added to the culture media.</p> <p>U2OS cells were washed twice with PBS (pH 7.4) and fixed for 30 min in 2% PFA in PBS, washed 3-times for 5 min with PBS, then permeabilized in 0.1% Triton X-100 in PBS for 20 min, washed 3-times for 5 min by PBS and blocked in filtered 5% BSA in PBS for 30 min. Cells were incubated for 45 min with rabbit polyclonal IgG anti-RNAPII CTD P-S2 (Abcam ab5095) 3 µg/mL and mouse monoclonal IgG2 anti-PI(3,4)P2 (Z-P034; Echelon Biosci. Inc., USA) 5 µg/mL in 5% BSA in PBS, washed 3-times for 5 min in PBS and incubated for 30 min with donkey anti-mouse IgG AF555 (Invitrogen A31570) 10 µg/mL and goat anti-rabbit IgG AF647 (Invitrogen A21245) 10 µg/mL diluted in 5% BSA in PBS. Then the cells were washed 3-times for 5 min in PBS, post-fixed for 15 min in 2% PFA in PBS and washed 3-times for 5 min in PBS. All procedures were performed at RT and the cells were stored in PBS in the fridge overnight prior imaging.</p> <p>Coverslips with cells were mounted in the Chamlide chamber (Live Cell Instrument, Korea) and covered with imaging buffer (PBS pH 7.4, 50 mM MEA). Single-molecule localizations (SMLs) data were acquired by Zeiss Elyra PS.1 equipped with HR Diode 642-150 and HR DPSS 561-200 lasers, Alpha Plan-Apochromat 100x/1.46 oil DIC M27 Elyra objective and Andor EM CCD iXon DU 897 camera and Zeiss ZEN Black 2.1 SP3 software (Zeiss). AF647 and AF555 photo-switching was achieved by HiLo illumination and TIRF HP FOV with 100% power of 642nm or 561nm laser, and the signal was acquired via MBS 642 + EF LP 655 and MBS 561 + EF BP 570-620 / LP 750 filters, respectively. Exposure time was 40 ms and EM gain was 300 for both channels.</p> <p>SMLs were calculated in 2D by Zeiss ZEN Black 2.1 SP3 software using x,y 2D Gauss fit with point spread function (PSF) half width 177.9 nm, peak mask size 9 pixels and peak intensity to noise 6 and accounted for overlap in 2D with max cluster size 10. SMLs were rendered in ZEN software with 10 nm/px resolution and 1x PSF expansion factor. The data were model-based drift corrected in ZEN. Two channels were aligned using tetraspec beads fiducial markers for affine calibration. Drift-corrected and aligned localization coordinates were exported as text files. Text files were converted into csv files and imported using self-written macro (Hoboth et al., 2021a) into the ImageJ2 (Rueden et al., 2017) plug-in ThunderSTORM, visualized by normalized Gaussian method (Ovesny et al., 2014).</p>
Ditrect STORM imaging and image reconstruction of the transcription elongation marker P-S2 and nuclear PI(3,4)P2 indirectly immunolabeled with AF647 (red) and AF555 (green) in control cells.
<p>U-2 OS cells were grown in DMEM with 10% FBS at 37°C and 5% CO<sub>2</sub>. Cells were plated one day before staining in ~50% confluence on the high-precision 12 mm round coverslips treated with Hellmanex, sonicated, washed, dried and sterilized. Cells were control treated with 1:000 DMSO in the culture media.</p> <p>U2OS cells were washed twice with PBS (pH 7.4) and fixed for 30 min in 2% PFA in PBS, washed 3-times for 5 min with PBS, then permeabilized in 0.1% Triton X-100 in PBS for 20 min, washed 3-times for 5 min by PBS and blocked in filtered 5% BSA in PBS for 30 min. Cells were incubated for 45 min with rabbit polyclonal IgG anti-RNAPII CTD P-S2 (Abcam ab5095) 3 µg/mL and mouse monoclonal IgG2 anti-PI(3,4)P2 (Z-P034; Echelon Biosci. Inc., USA) 5 µg/mL in 5% BSA in PBS, washed 3-times for 5 min in PBS and incubated for 30 min with donkey anti-mouse IgG AF555 (Invitrogen A31570) 10 µg/mL and goat anti-rabbit IgG AF647 (Invitrogen A21245) 10 µg/mL diluted in 5% BSA in PBS. Then the cells were washed 3-times for 5 min in PBS, post-fixed for 15 min in 2% PFA in PBS and washed 3-times for 5 min in PBS. All procedures were performed at RT and the cells were stored in PBS in the fridge overnight prior imaging.</p> <p>Coverslips with cells were mounted in the Chamlide chamber (Live Cell Instrument, Korea) and covered with imaging buffer (PBS pH 7.4, 50 mM MEA). Single-molecule localizations (SMLs) data were acquired by Zeiss Elyra PS.1 equipped with HR Diode 642-150 and HR DPSS 561-200 lasers, Alpha Plan-Apochromat 100x/1.46 oil DIC M27 Elyra objective and Andor EM CCD iXon DU 897 camera and Zeiss ZEN Black 2.1 SP3 software (Zeiss). AF647 and AF555 photo-switching was achieved by HiLo illumination and TIRF HP FOV with 100% power of 642nm or 561nm laser, and the signal was acquired via MBS 642 + EF LP 655 and MBS 561 + EF BP 570-620 / LP 750 filters, respectively. Exposure time was 40 ms and EM gain was 300 for both channels.</p> <p>SMLs were calculated in 2D by Zeiss ZEN Black 2.1 SP3 software using x,y 2D Gauss fit with point spread function (PSF) half width 177.9 nm, peak mask size 9 pixels and peak intensity to noise 6 and accounted for overlap in 2D with max cluster size 10. SMLs were rendered in ZEN software with 10 nm/px resolution and 1x PSF expansion factor. The data were model-based drift corrected in ZEN. Two channels were aligned using tetraspec beads fiducial markers for affine calibration. Drift-corrected and aligned localization coordinates were exported as text files. Text files were converted into csv files and imported using self-written macro (Hoboth et al., 2021a) into the ImageJ2 (Rueden et al., 2017) plug-in ThunderSTORM, visualized by normalized Gaussian method (Ovesny et al., 2014).</p>
Ditrect STORM imaging and image reconstruction of the transcription initiation marker P-S2 and nuclear PI(3,4)P2 indirectly immunolabeled with AF647 (red) and AF555 (green) in DRB treated cells.
<p>U-2 OS cells were grown in DMEM with 10% FBS at 37°C and 5% CO<sub>2</sub>. Cells were plated one day before staining in ~50% confluence on the high-precision 12 mm round coverslips treated with Hellmanex, sonicated, washed, dried and sterilized. Cells were treated for 2h with 100 µM DRB (Sigma D1916) added to the cell culture media.</p> <p>U2OS cells were washed twice with PBS (pH 7.4) and fixed for 30 min in 2% PFA in PBS, washed 3-times for 5 min with PBS, then permeabilized in 0.1% Triton X-100 in PBS for 20 min, washed 3-times for 5 min by PBS and blocked in filtered 5% BSA in PBS for 30 min. Cells were incubated for 45 min with rabbit polyclonal IgG anti-RNAPII CTD P-S2 (Abcam ab5095) 3 µg/mL and mouse monoclonal IgG2 anti-PI(3,4)P2 (Z-P034; Echelon Biosci. Inc., USA) 5 µg/mL in 5% BSA in PBS, washed 3-times for 5 min in PBS and incubated for 30 min with donkey anti-mouse IgG AF555 (Invitrogen A31570) 10 µg/mL and goat anti-rabbit IgG AF647 (Invitrogen A21245) 10 µg/mL diluted in 5% BSA in PBS. Then the cells were washed 3-times for 5 min in PBS, post-fixed for 15 min in 2% PFA in PBS and washed 3-times for 5 min in PBS. All procedures were performed at RT and the cells were stored in PBS in the fridge overnight prior imaging.</p> <p>Coverslips with cells were mounted in the Chamlide chamber (Live Cell Instrument, Korea) and covered with imaging buffer (PBS pH 7.4, 50 mM MEA). Single-molecule localizations (SMLs) data were acquired by Zeiss Elyra PS.1 equipped with HR Diode 642-150 and HR DPSS 561-200 lasers, Alpha Plan-Apochromat 100x/1.46 oil DIC M27 Elyra objective and Andor EM CCD iXon DU 897 camera and Zeiss ZEN Black 2.1 SP3 software (Zeiss). AF647 and AF555 photo-switching was achieved by HiLo illumination and TIRF HP FOV with 100% power of 642nm or 561nm laser, and the signal was acquired via MBS 642 + EF LP 655 and MBS 561 + EF BP 570-620 / LP 750 filters, respectively. Exposure time was 40 ms and EM gain was 300 for both channels.</p> <p>SMLs were calculated in 2D by Zeiss ZEN Black 2.1 SP3 software using x,y 2D Gauss fit with point spread function (PSF) half width 177.9 nm, peak mask size 9 pixels and peak intensity to noise 6 and accounted for overlap in 2D with max cluster size 10. SMLs were rendered in ZEN software with 10 nm/px resolution and 1x PSF expansion factor. The data were model-based drift corrected in ZEN. Two channels were aligned using tetraspec beads fiducial markers for affine calibration. Drift-corrected and aligned localization coordinates were exported as text files. Text files were converted into csv files and imported using self-written macro (Hoboth et al., 2021a) into the ImageJ2 (Rueden et al., 2017) plug-in ThunderSTORM, visualized by normalized Gaussian method (Ovesny et al., 2014).</p>
Ditrect STORM imaging and image reconstruction of the transcription initiation marker P-S5 and nuclear PI(4,5)P2 indirectly immunolabeled with AF647 (red) and AF555 (green) in THZ1 treated cells.
<p>U-2 OS cells were grown in DMEM with 10% FBS at 37°C and 5% CO<sub>2</sub>. Cells were plated one day before staining in ~50% confluence on the high-precision 12 mm round coverslips treated with Hellmanex, sonicated, washed, dried and sterilized. Cells were treated for 3h with 1 µM THZ1 (MedChem HY80013) added to the culture media.</p> <p>U2OS cells were washed twice with PBS (pH 7.4) and fixed for 30 min in 2% PFA in PBS, washed 3-times for 5 min with PBS, then permeabilized in 0.1% Triton X-100 in PBS for 20 min, washed 3-times for 5 min by PBS and blocked in filtered 5% BSA in PBS for 30 min. Cells were incubated for 45 min with rabbit polyclonal IgG anti-RNAPII CTD P-S5 (Abcam ab5131) 3 µg/mL and mouse ascites IgM anti-PI(4,5)P2 2C11 (Z-A045; Echelon Biosci. Inc., USA) 5 µg/mL in 5% BSA in PBS, washed 3-times for 5 min in PBS and incubated for 30 min with goat anti-mouse IgM (µ-chain) AF555 (Jackson ImmunoRes. A24126) 10 µg/mL; goat anti-rabbit IgG AF647 (Invitrogen A21245) 10 µg/mL diluted in 5% BSA in PBS. Then the cells were washed 3-times for 5 min in PBS, post-fixed for 15 min in 2% PFA in PBS and washed 3-times for 5 min in PBS. All procedures were performed at RT and the cells were stored in PBS in the fridge overnight prior imaging.</p> <p>Coverslips with cells were mounted in the Chamlide chamber (Live Cell Instrument, Korea) and covered with imaging buffer (PBS pH 7.4, 50 mM MEA). Single-molecule localizations (SMLs) data were acquired by Zeiss Elyra PS.1 equipped with HR Diode 642-150 and HR DPSS 561-200 lasers, Alpha Plan-Apochromat 100x/1.46 oil DIC M27 Elyra objective and Andor EM CCD iXon DU 897 camera and Zeiss ZEN Black 2.1 SP3 software (Zeiss). AF647 and AF555 photo-switching was achieved by HiLo illumination and TIRF HP FOV with 100% power of 642nm or 561nm laser, and the signal was acquired via MBS 642 + EF LP 655 and MBS 561 + EF BP 570-620 / LP 750 filters, respectively. Exposure time was 40 ms and EM gain was 300 for both channels.</p> <p>SMLs were calculated in 2D by Zeiss ZEN Black 2.1 SP3 software using x,y 2D Gauss fit with point spread function (PSF) half width 177.9 nm, peak mask size 9 pixels and peak intensity to noise 6 and accounted for overlap in 2D with max cluster size 10. SMLs were rendered in ZEN software with 10 nm/px resolution and 1x PSF expansion factor. The data were model-based drift corrected in ZEN. Two channels were aligned using tetraspec beads fiducial markers for affine calibration. Drift-corrected and aligned localization coordinates were exported as text files. Text files were converted into csv files and imported using self-written macro (Hoboth et al., 2021a) into the ImageJ2 (Rueden et al., 2017) plug-in ThunderSTORM, visualized by normalized Gaussian method (Ovesny et al., 2014).</p>
Ditrect STORM imaging and image reconstruction of the transcription initiation marker P-S5 and nuclear PI(3,4)P2 indirectly immunolabeled with AF647 (red) and AF555 (green) in THZ1 treated cells.
<p>U-2 OS cells were grown in DMEM with 10% FBS at 37°C and 5% CO<sub>2</sub>. Cells were plated one day before staining in ~50% confluence on the high-precision 12 mm round coverslips treated with Hellmanex, sonicated, washed, dried and sterilized. Cells were treated for 3h with 1 µM THZ1 (MedChem HY80013) added to the culture media.</p> <p>U2OS cells were washed twice with PBS (pH 7.4) and fixed for 30 min in 2% PFA in PBS, washed 3-times for 5 min with PBS, then permeabilized in 0.1% Triton X-100 in PBS for 20 min, washed 3-times for 5 min by PBS and blocked in filtered 5% BSA in PBS for 30 min. Cells were incubated for 45 min with rabbit polyclonal IgG anti-RNAPII CTD P-S5 (Abcam ab5131) 3 µg/mL and mouse monoclonal IgG2 anti-PI(3,4)P2 (Z-P034; Echelon Biosci. Inc., USA) 5 µg/mL in 5% BSA in PBS, washed 3-times for 5 min in PBS and incubated for 30 min with donkey anti-mouse IgG AF555 (Invitrogen A31570) 10 µg/mL and goat anti-rabbit IgG AF647 (Invitrogen A21245) 10 µg/mL diluted in 5% BSA in PBS. Then the cells were washed 3-times for 5 min in PBS, post-fixed for 15 min in 2% PFA in PBS and washed 3-times for 5 min in PBS. All procedures were performed at RT and the cells were stored in PBS in the fridge overnight prior imaging.</p> <p>Coverslips with cells were mounted in the Chamlide chamber (Live Cell Instrument, Korea) and covered with imaging buffer (PBS pH 7.4, 50 mM MEA). Single-molecule localizations (SMLs) data were acquired by Zeiss Elyra PS.1 equipped with HR Diode 642-150 and HR DPSS 561-200 lasers, Alpha Plan-Apochromat 100x/1.46 oil DIC M27 Elyra objective and Andor EM CCD iXon DU 897 camera and Zeiss ZEN Black 2.1 SP3 software (Zeiss). AF647 and AF555 photo-switching was achieved by HiLo illumination and TIRF HP FOV with 100% power of 642nm or 561nm laser, and the signal was acquired via MBS 642 + EF LP 655 and MBS 561 + EF BP 570-620 / LP 750 filters, respectively. Exposure time was 40 ms and EM gain was 300 for both channels.</p> <p>SMLs were calculated in 2D by Zeiss ZEN Black 2.1 SP3 software using x,y 2D Gauss fit with point spread function (PSF) half width 177.9 nm, peak mask size 9 pixels and peak intensity to noise 6 and accounted for overlap in 2D with max cluster size 10. SMLs were rendered in ZEN software with 10 nm/px resolution and 1x PSF expansion factor. The data were model-based drift corrected in ZEN. Two channels were aligned using tetraspec beads fiducial markers for affine calibration. Drift-corrected and aligned localization coordinates were exported as text files. Text files were converted into csv files and imported using self-written macro (Hoboth et al., 2021a) into the ImageJ2 (Rueden et al., 2017) plug-in ThunderSTORM, visualized by normalized Gaussian method (Ovesny et al., 2014).</p>
Ditrect STORM imaging and image reconstruction of the transcription initiation marker P-S5 and nuclear PI(3,4)P2 indirectly immunolabeled with AF647 (red) and AF555 (green) in DRB treated cells.
<p>U-2 OS cells were grown in DMEM with 10% FBS at 37°C and 5% CO<sub>2</sub>. Cells were plated one day before staining in ~50% confluence on the high-precision 12 mm round coverslips treated with Hellmanex, sonicated, washed, dried and sterilized. Cells were treated for 2h with 100 µM DRB (Sigma D1916) added to the cell culture media.</p> <p>U2OS cells were washed twice with PBS (pH 7.4) and fixed for 30 min in 2% PFA in PBS, washed 3-times for 5 min with PBS, then permeabilized in 0.1% Triton X-100 in PBS for 20 min, washed 3-times for 5 min by PBS and blocked in filtered 5% BSA in PBS for 30 min. Cells were incubated for 45 min with rabbit polyclonal IgG anti-RNAPII CTD P-S5 (Abcam ab5131) 3 µg/mL and mouse monoclonal IgG2 anti-PI(3,4)P2 (Z-P034; Echelon Biosci. Inc., USA) 5 µg/mL in 5% BSA in PBS, washed 3-times for 5 min in PBS and incubated for 30 min with donkey anti-mouse IgG AF555 (Invitrogen A31570) 10 µg/mL and goat anti-rabbit IgG AF647 (Invitrogen A21245) 10 µg/mL diluted in 5% BSA in PBS. Then the cells were washed 3-times for 5 min in PBS, post-fixed for 15 min in 2% PFA in PBS and washed 3-times for 5 min in PBS. All procedures were performed at RT and the cells were stored in PBS in the fridge overnight prior imaging.</p> <p>Coverslips with cells were mounted in the Chamlide chamber (Live Cell Instrument, Korea) and covered with imaging buffer (PBS pH 7.4, 50 mM MEA). Single-molecule localizations (SMLs) data were acquired by Zeiss Elyra PS.1 equipped with HR Diode 642-150 and HR DPSS 561-200 lasers, Alpha Plan-Apochromat 100x/1.46 oil DIC M27 Elyra objective and Andor EM CCD iXon DU 897 camera and Zeiss ZEN Black 2.1 SP3 software (Zeiss). AF647 and AF555 photo-switching was achieved by HiLo illumination and TIRF HP FOV with 100% power of 642nm or 561nm laser, and the signal was acquired via MBS 642 + EF LP 655 and MBS 561 + EF BP 570-620 / LP 750 filters, respectively. Exposure time was 40 ms and EM gain was 300 for both channels.</p> <p>SMLs were calculated in 2D by Zeiss ZEN Black 2.1 SP3 software using x,y 2D Gauss fit with point spread function (PSF) half width 177.9 nm, peak mask size 9 pixels and peak intensity to noise 6 and accounted for overlap in 2D with max cluster size 10. SMLs were rendered in ZEN software with 10 nm/px resolution and 1x PSF expansion factor. The data were model-based drift corrected in ZEN. Two channels were aligned using tetraspec beads fiducial markers for affine calibration. Drift-corrected and aligned localization coordinates were exported as text files. Text files were converted into csv files and imported using self-written macro (Hoboth et al., 2021a) into the ImageJ2 (Rueden et al., 2017) plug-in ThunderSTORM, visualized by normalized Gaussian method (Ovesny et al., 2014).</p>
Ditrect STORM imaging and image reconstruction of the transcription elongation marker P-S2 and nuclear PI(4,5)P2 indirectly immunolabeled with AF647 (red) and AF555 (green) in control cells.
<p>U-2 OS cells were grown in DMEM with 10% FBS at 37°C and 5% CO<sub>2</sub>. Cells were plated one day before staining in ~50% confluence on the high-precision 12 mm round coverslips treated with Hellmanex, sonicated, washed, dried and sterilized. Cells were control treated with 1:000 DMSO in the culture media.</p> <p>U2OS cells were washed twice with PBS (pH 7.4) and fixed for 30 min in 2% PFA in PBS, washed 3-times for 5 min with PBS, then permeabilized in 0.1% Triton X-100 in PBS for 20 min, washed 3-times for 5 min by PBS and blocked in filtered 5% BSA in PBS for 30 min. Cells were incubated for 45 min with rabbit polyclonal IgG anti-RNAPII CTD P-S2 (Abcam ab5095) 3 µg/mL and mouse ascites IgM anti-PI(4,5)P2 2C11 (Z-A045; Echelon Biosci. Inc., USA) 5 µg/mL in 5% BSA in PBS, washed 3-times for 5 min in PBS and incubated for 30 min with goat anti-mouse IgM (µ-chain) AF555 (Jackson ImmunoRes. A24126) 10 µg/mL; goat anti-rabbit IgG AF647 (Invitrogen A21245) 10 µg/mL diluted in 5% BSA in PBS. Then the cells were washed 3-times for 5 min in PBS, post-fixed for 15 min in 2% PFA in PBS and washed 3-times for 5 min in PBS. All procedures were performed at RT and the cells were stored in PBS in the fridge overnight prior imaging.</p> <p>Coverslips with cells were mounted in the Chamlide chamber (Live Cell Instrument, Korea) and covered with imaging buffer (PBS pH 7.4, 50 mM MEA). Single-molecule localizations (SMLs) data were acquired by Zeiss Elyra PS.1 equipped with HR Diode 642-150 and HR DPSS 561-200 lasers, Alpha Plan-Apochromat 100x/1.46 oil DIC M27 Elyra objective and Andor EM CCD iXon DU 897 camera and Zeiss ZEN Black 2.1 SP3 software (Zeiss). AF647 and AF555 photo-switching was achieved by HiLo illumination and TIRF HP FOV with 100% power of 642nm or 561nm laser, and the signal was acquired via MBS 642 + EF LP 655 and MBS 561 + EF BP 570-620 / LP 750 filters, respectively. Exposure time was 40 ms and EM gain was 300 for both channels.</p> <p>SMLs were calculated in 2D by Zeiss ZEN Black 2.1 SP3 software using x,y 2D Gauss fit with point spread function (PSF) half width 177.9 nm, peak mask size 9 pixels and peak intensity to noise 6 and accounted for overlap in 2D with max cluster size 10. SMLs were rendered in ZEN software with 10 nm/px resolution and 1x PSF expansion factor. The data were model-based drift corrected in ZEN. Two channels were aligned using tetraspec beads fiducial markers for affine calibration. Drift-corrected and aligned localization coordinates were exported as text files. Text files were converted into csv files and imported using self-written macro (Hoboth et al., 2021a) into the ImageJ2 (Rueden et al., 2017) plug-in ThunderSTORM, visualized by normalized Gaussian method (Ovesny et al., 2014).</p>
Ditrect STORM imaging and image reconstruction of the transcription elongation marker P-S2 and nuclear PI(4,5)P2 indirectly immunolabeled with AF647 (red) and AF555 (green) in DRB treated cells.
<p>U-2 OS cells were grown in DMEM with 10% FBS at 37°C and 5% CO<sub>2</sub>. Cells were plated one day before staining in ~50% confluence on the high-precision 12 mm round coverslips treated with Hellmanex, sonicated, washed, dried and sterilized. Cells were treated for 2h with 100 µM DRB (Sigma D1916) added to the cell culture media.</p> <p>U2OS cells were washed twice with PBS (pH 7.4) and fixed for 30 min in 2% PFA in PBS, washed 3-times for 5 min with PBS, then permeabilized in 0.1% Triton X-100 in PBS for 20 min, washed 3-times for 5 min by PBS and blocked in filtered 5% BSA in PBS for 30 min. Cells were incubated for 45 min with rabbit polyclonal IgG anti-RNAPII CTD P-S2 (Abcam ab5095) 3 µg/mL and mouse ascites IgM anti-PI(4,5)P2 2C11 (Z-A045; Echelon Biosci. Inc., USA) 5 µg/mL in 5% BSA in PBS, washed 3-times for 5 min in PBS and incubated for 30 min with goat anti-mouse IgM (µ-chain) AF555 (Jackson ImmunoRes. A24126) 10 µg/mL; goat anti-rabbit IgG AF647 (Invitrogen A21245) 10 µg/mL diluted in 5% BSA in PBS. Then the cells were washed 3-times for 5 min in PBS, post-fixed for 15 min in 2% PFA in PBS and washed 3-times for 5 min in PBS. All procedures were performed at RT and the cells were stored in PBS in the fridge overnight prior imaging.</p> <p>Coverslips with cells were mounted in the Chamlide chamber (Live Cell Instrument, Korea) and covered with imaging buffer (PBS pH 7.4, 50 mM MEA). Single-molecule localizations (SMLs) data were acquired by Zeiss Elyra PS.1 equipped with HR Diode 642-150 and HR DPSS 561-200 lasers, Alpha Plan-Apochromat 100x/1.46 oil DIC M27 Elyra objective and Andor EM CCD iXon DU 897 camera and Zeiss ZEN Black 2.1 SP3 software (Zeiss). AF647 and AF555 photo-switching was achieved by HiLo illumination and TIRF HP FOV with 100% power of 642nm or 561nm laser, and the signal was acquired via MBS 642 + EF LP 655 and MBS 561 + EF BP 570-620 / LP 750 filters, respectively. Exposure time was 40 ms and EM gain was 300 for both channels.</p> <p>SMLs were calculated in 2D by Zeiss ZEN Black 2.1 SP3 software using x,y 2D Gauss fit with point spread function (PSF) half width 177.9 nm, peak mask size 9 pixels and peak intensity to noise 6 and accounted for overlap in 2D with max cluster size 10. SMLs were rendered in ZEN software with 10 nm/px resolution and 1x PSF expansion factor. The data were model-based drift corrected in ZEN. Two channels were aligned using tetraspec beads fiducial markers for affine calibration. Drift-corrected and aligned localization coordinates were exported as text files. Text files were converted into csv files and imported using self-written macro (Hoboth et al., 2021a) into the ImageJ2 (Rueden et al., 2017) plug-in ThunderSTORM, visualized by normalized Gaussian method (Ovesny et al., 2014).</p>
Ditrect STORM imaging and image reconstruction of the transcription initiation marker P-S5 and nuclear PI(3,4)P2 indirectly immunolabeled with AF647 (red) and AF555 (green) in control cells.
<p>U-2 OS cells were grown in DMEM with 10% FBS at 37°C and 5% CO<sub>2</sub>. Cells were plated one day before staining in ~50% confluence on the high-precision 12 mm round coverslips treated with Hellmanex, sonicated, washed, dried and sterilized. Cells were control treated with 1:000 DMSO in the culture media.</p> <p>U2OS cells were washed twice with PBS (pH 7.4) and fixed for 30 min in 2% PFA in PBS, washed 3-times for 5 min with PBS, then permeabilized in 0.1% Triton X-100 in PBS for 20 min, washed 3-times for 5 min by PBS and blocked in filtered 5% BSA in PBS for 30 min. Cells were incubated for 45 min with rabbit polyclonal IgG anti-RNAPII CTD P-S5 (Abcam ab5131) 3 µg/mL and mouse monoclonal IgG2 anti-PI(3,4)P2 (Z-P034; Echelon Biosci. Inc., USA) 5 µg/mL in 5% BSA in PBS, washed 3-times for 5 min in PBS and incubated for 30 min with donkey anti-mouse IgG AF555 (Invitrogen A31570) 10 µg/mL and goat anti-rabbit IgG AF647 (Invitrogen A21245) 10 µg/mL diluted in 5% BSA in PBS. Then the cells were washed 3-times for 5 min in PBS, post-fixed for 15 min in 2% PFA in PBS and washed 3-times for 5 min in PBS. All procedures were performed at RT and the cells were stored in PBS in the fridge overnight prior imaging.</p> <p>Coverslips with cells were mounted in the Chamlide chamber (Live Cell Instrument, Korea) and covered with imaging buffer (PBS pH 7.4, 50 mM MEA). Single-molecule localizations (SMLs) data were acquired by Zeiss Elyra PS.1 equipped with HR Diode 642-150 and HR DPSS 561-200 lasers, Alpha Plan-Apochromat 100x/1.46 oil DIC M27 Elyra objective and Andor EM CCD iXon DU 897 camera and Zeiss ZEN Black 2.1 SP3 software (Zeiss). AF647 and AF555 photo-switching was achieved by HiLo illumination and TIRF HP FOV with 100% power of 642nm or 561nm laser, and the signal was acquired via MBS 642 + EF LP 655 and MBS 561 + EF BP 570-620 / LP 750 filters, respectively. Exposure time was 40 ms and EM gain was 300 for both channels.</p> <p>SMLs were calculated in 2D by Zeiss ZEN Black 2.1 SP3 software using x,y 2D Gauss fit with point spread function (PSF) half width 177.9 nm, peak mask size 9 pixels and peak intensity to noise 6 and accounted for overlap in 2D with max cluster size 10. SMLs were rendered in ZEN software with 10 nm/px resolution and 1x PSF expansion factor. The data were model-based drift corrected in ZEN. Two channels were aligned using tetraspec beads fiducial markers for affine calibration. Drift-corrected and aligned localization coordinates were exported as text files. Text files were converted into csv files and imported using self-written macro (Hoboth et al., 2021a) into the ImageJ2 (Rueden et al., 2017) plug-in ThunderSTORM, visualized by normalized Gaussian method (Ovesny et al., 2014).</p>
SMART: Spatial transcriptomics deconvolution using marker-gene-assisted topic model
<p>Source code and simulated datasets used in manuscript "SMART: Spatial transcriptomics deconvolution using marker-gene-assisted topic model"</p>
Appendix 2 in A new subfamily classification of the Citrus family (Rutaceae) based on six nuclear and plastid markers
<p><b>Appendix 2.</b> Details about the character states of all included taxa for the 13 morphological and karyological characters.</p><table><thead><tr><th></th><th></th><th></th><th></th><th></th><th></th><th colspan="2">Characters</th><th></th><th></th><th></th><th></th><th></th><th></th></tr></thead><tbody><tr><th>Taxon</th><td>1</td><td>2</td><td>3</td><td>4</td><td>5</td><td colspan="2">6 7</td><td>8</td><td>9</td><td>10</td><td>11</td><td>12</td><td>13</td></tr><tr><th><i>Acmadenia</i></th><td>0</td><td>0</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Acradenia</i></th><td>1</td><td>1</td><td>2, 3</td><td>1</td><td>1</td><td>1</td><td>2</td><td>1</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Acronychia</i></th><td>0, 1</td><td>1</td><td>1</td><td>1</td><td>0, 1</td><td>1</td><td>1</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Adenandra</i></th><td>0</td><td>0, 1</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>0</td><td>2, 3</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Adiscanthus</i></th><td>0</td><td>0</td><td>2</td><td>0</td><td>0</td><td>1</td><td>2</td><td>1</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Aegle</i></th><td>0, 1</td><td>0</td><td>1, 2</td><td>2</td><td>0</td><td>4</td><td>0</td><td>0</td><td>0, 1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Aeglopsis</i></th><td>0, 1</td><td>0</td><td>1, 2, 3</td><td>1, 2</td><td>0</td><td>4</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Afraegle</i></th><td>1</td><td>0</td><td>1, 2, 3</td><td>2</td><td>0</td><td>4</td><td>0</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Agathosma</i> A</th><td>0</td><td>0, 1, 2</td><td>2, 3</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Agathosma</i> B</th><td>0</td><td>0, 1, 2</td><td>2, 3</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Agathosma</i> C</th><td>0</td><td>0, 1, 2</td><td>2, 3</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Amyris</i></th><td>0, 1</td><td>0, 1</td><td>1, 2, 3</td><td>1</td><td>3</td><td>1</td><td>1</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Andreadoxa</i></th><td>0</td><td>0</td><td>2</td><td>0</td><td>1</td><td>1</td><td>2</td><td>1</td><td>?</td><td>0</td><td>1</td><td>0</td><td>0</td></tr><tr><th><i>Angostura</i></th><td>0, 1</td><td>0</td><td>2</td><td>0</td><td>1</td><td>1</td><td>2</td><td>1</td><td>?</td><td>0</td><td>0, 1</td><td>1</td><td>0</td></tr><tr><th><i>Asterolasia</i></th><td>0</td><td>0</td><td>2</td><td>1</td><td>0, 1</td><td>0, 1</td><td>2</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Atalantia</i></th><td>0</td><td>0</td><td>1, 2, 3</td><td>1</td><td>0</td><td>0, 1</td><td>5</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Balfourodendron</i></th><td>1</td><td>1</td><td>1</td><td>0</td><td>0</td><td>1</td><td>4</td><td>1</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Balsamocitrus</i></th><td>0, 1</td><td>0</td><td>2</td><td>1</td><td>0</td><td>4</td><td>0</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Bergera</i></th><td>0, 1</td><td>0</td><td>1, 2</td><td>1</td><td>0</td><td>?</td><td>0</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Boenninghausenia</i></th><td>1</td><td>0</td><td>1</td><td>1</td><td>1</td><td>2, 3</td><td>2</td><td>2</td><td>0</td><td>2</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Boronia</i> A</th><td>0, 1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>0, 1, 2</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Boronia</i> B</th><td>0, 1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>0, 1, 2</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Boronia</i> C</th><td>0, 1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>0, 1, 2</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Boronia</i> D (<i>Boronella</i>)</th><td>0</td><td>1, 2</td><td>1</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Bosistoa</i></th><td>0, 1</td><td>0</td><td>2</td><td>1</td><td>1</td><td>2, 3</td><td>2</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Bottegoa</i></th><td>1</td><td>0</td><td>1, 2</td><td>0</td><td>0</td><td>0</td><td>4</td><td>1</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Bouchardatia</i></th><td>0, 1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>3</td><td>2</td><td>0, 1</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Brombya</i></th><td>0</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>2</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Burkillanthus</i></th><td>0, 1</td><td>0</td><td>2</td><td>1</td><td>0</td><td>4</td><td>5</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Calodendrum</i></th><td>0</td><td>1, 2</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>0</td><td>2</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Casimiroa</i></th><td>0, 1</td><td>0</td><td>1, 2, 3</td><td>0</td><td>0</td><td>1, 2, 3</td><td>0, 1</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Cedrelopsis</i></th><td>1</td><td>0</td><td>2</td><td>0</td><td>0</td><td>1, 2</td><td>2</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>1</td></tr><tr><th><i>Chloroxylon</i></th><td>1</td><td>0</td><td>2</td><td>1</td><td>0</td><td>3</td><td>2</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1</td></tr><tr><th><i>Choisya</i></th><td>1</td><td>1</td><td>1, 2</td><td>1</td><td>1</td><td>1</td><td>2</td><td>2</td><td>2</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Chorilaena</i></th><td>0</td><td>0</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Citropsis</i></th><td>0, 1</td><td>0</td><td>1, 2</td><td>1</td><td>0</td><td>0</td><td>5</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Citrus</i> A</th><td>0</td><td>0</td><td>1, 2, 3</td><td>2</td><td>0</td><td>1, 2, 3</td><td>5</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Citrus</i> B</th><td>0</td><td>0</td><td>1, 2, 3</td><td>2</td><td>0</td><td>1, 2, 3</td><td>5</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th>Taxon</th><td>1</td><td>2</td><td>3</td><td>4</td><td>5</td><td colspan="2">Characters 6 7</td><td>8</td><td>9</td><td>10</td><td>11</td><td>12</td><td>13</td></tr><tr><th><i>Citrus</i> C</th><td>0</td><td>0</td><td>1, 2, 3</td><td>2</td><td>0</td><td>1, 2, 3</td><td>5</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Clausena</i></th><td>1</td><td>0</td><td>1, 2</td><td>1</td><td>0</td><td>1</td><td>0</td><td>0</td><td>0, 1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Clymenia</i> (<i>Citrus</i> s.l.)</th><td>0</td><td>0</td><td>1, 2, 3</td><td>2</td><td>0</td><td>1, 2, 3</td><td>5</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Cneoridium</i></th><td>0</td><td>1</td><td>1</td><td>1</td><td>3</td><td>1, 2</td><td>0</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Cneorum</i></th><td>0</td><td>0</td><td>0, 1</td><td>0</td><td>0</td><td>1</td><td>1</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Coatesia</i></th><td>0</td><td>0</td><td>2</td><td>0</td><td>1</td><td>1</td><td>2</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Coleonema</i></th><td>0</td><td>0</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>0</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Comptonella</i></th><td>0, 1</td><td>1</td><td>1</td><td>0, 1</td><td>0, 1</td><td>1</td><td>1</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Conchocarpus</i> A</th><td>0, 1</td><td>0, 1</td><td>1, 2</td><td>0</td><td>1</td><td>1</td><td>2</td><td>1</td><td>?</td><td>0</td><td>0, 1</td><td>1</td><td>0</td></tr><tr><th><i>Conchocarpus</i> B (<i>Almeidea</i>)</th><td>0</td><td>0</td><td>2</td><td>0</td><td>1</td><td>1</td><td>2</td><td>1</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Correa</i></th><td>0</td><td>1</td><td>1</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>1</td><td>0</td><td>0</td><td>1</td><td>0</td></tr><tr><th><i>Crossosperma</i></th><td>1</td><td>1</td><td>1</td><td>1</td><td>0</td><td>0, 1</td><td>1</td><td>1</td><td>?</td><td>0</td><td>0</td><td>0</td><td>1</td></tr><tr><th><i>Crowea</i></th><td>0</td><td>0</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Cyanothamnus</i></th><td>0, 1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Desmotes</i></th><td>0</td><td>1</td><td>2</td><td>0</td><td>1</td><td>1</td><td>2</td><td>1</td><td>?</td><td>0</td><td>1</td><td>1</td><td>0</td></tr><tr><th><i>Dictamnus</i></th><td>1</td><td>0</td><td>2</td><td>1</td><td>0</td><td>2</td><td>2</td><td>2</td><td>1</td><td>2</td><td>1</td><td>0</td><td>0</td></tr><tr><th><i>Dictyoloma</i></th><td>1</td><td>0</td><td>2</td><td>0</td><td>1</td><td>2</td><td>2</td><td>1</td><td>?</td><td>0</td><td>0</td><td>0</td><td>1</td></tr><tr><th><i>Dinosperma</i></th><td>0, 1</td><td>0, 1, 2</td><td>1</td><td>1</td><td>1</td><td>1</td><td>2</td><td>0, 1</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Diosma</i></th><td>0</td><td>0, 1</td><td>2</td><td>0, 1</td><td>1</td><td>0, 1</td><td>2</td><td>0</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Diplolaena</i></th><td>0</td><td>0</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Drummondita</i></th><td>0</td><td>0</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Dryades</i></th><td>0</td><td>0</td><td>2</td><td>0</td><td>1</td><td>1</td><td>2</td><td>?</td><td>?</td><td>0</td><td>0, 1</td><td>1</td><td>0</td></tr><tr><th><i>Dutaillyea</i></th><td>0, 1</td><td>1</td><td>1</td><td>1</td><td>0</td><td>0, 1</td><td>1</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Empleurum</i></th><td>0</td><td>0</td><td>1</td><td>0</td><td>1, 3</td><td>0, 1</td><td>2</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Eremocitrus</i> (<i>Citrus</i> s.l.)</th><td>0</td><td>0</td><td>1, 2, 3</td><td>2</td><td>0</td><td>1, 2, 3</td><td>5</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Eriostemon</i></th><td>0</td><td>0</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Ertela</i></th><td>1</td><td>0, 1</td><td>2</td><td>0</td><td>1</td><td>1</td><td>2</td><td>1</td><td>?</td><td>1, 2</td><td>1</td><td>1</td><td>0</td></tr><tr><th><i>Erythrochiton</i></th><td>0</td><td>0</td><td>2</td><td>0</td><td>1</td><td>1</td><td>2</td><td>1</td><td>?</td><td>0</td><td>0, 1</td><td>1</td><td>0</td></tr><tr><th><i>Esenbeckia</i></th><td>0, 1</td><td>0, 1</td><td>2, 3</td><td>0</td><td>0, 1</td><td>1</td><td>2</td><td>0, 1</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Euchaetis</i></th><td>0</td><td>0, 1</td><td>2</td><td>0, 1</td><td>1</td><td>0, 1</td><td>2</td><td>0</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Euodia</i></th><td>0, 1</td><td>1</td><td>1</td><td>0, 1</td><td>1</td><td>1</td><td>2</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Fagaropsis</i></th><td>1</td><td>1</td><td>1, 2, 3</td><td>0, 1</td><td>0</td><td>0</td><td>1</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Feroniella</i></th><td>1</td><td>0</td><td>1, 2, 3</td><td>2</td><td>0</td><td>1, 2, 3</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Flindersia</i></th><td>0, 1</td><td>0, 1</td><td>2</td><td>1</td><td>0</td><td>1, 2, 3</td><td>2</td><td>0</td><td>1, 3</td><td>0</td><td>0</td><td>0</td><td>1</td></tr><tr><th><i>Fortunella</i> (<i>Citrus</i> s.l.)</th><td>0</td><td>0</td><td>1, 2, 3</td><td>2</td><td>0</td><td>1, 2, 3</td><td>5</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Galipea</i></th><td>0, 1</td><td>0</td><td>2</td><td>0</td><td>0</td><td>1</td><td>2</td><td>1</td><td>?</td><td>0</td><td>1</td><td>1</td><td>0</td></tr><tr><th><i>Geijera</i></th><td>0</td><td>0</td><td>0, 1, 2</td><td>0</td><td>1</td><td>1</td><td>3</td><td>1</td><td>3</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Geleznowia</i></th><td>0</td><td>0</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Glycosmis</i></th><td>0, 1</td><td>0</td><td>1, 2</td><td>0, 1</td><td>0</td><td>0, 1</td><td>0</td><td>0</td><td>0, 2</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Halfordia</i></th><td>0</td><td>0</td><td>2</td><td>1</td><td>0</td><td>0</td><td>1</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Haplophyllum</i></th><td>0, 1</td><td>0</td><td>2</td><td>1</td><td>1</td><td>0, 1, 2, 3</td><td>2</td><td>2</td><td>0</td><td>1, 2</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Harrisonia</i></th><td>1</td><td>0</td><td>1, 2</td><td>1</td><td>0</td><td>0</td><td>1</td><td>1</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Helietta</i></th><td>1</td><td>0, 1</td><td>1, 2</td><td>0</td><td>0</td><td>1</td><td>4</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Hortia</i></th><td>0</td><td>0</td><td>2</td><td>0</td><td>0</td><td>1</td><td>0, 1</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th>Taxon</th><td>1</td><td>2</td><td>3</td><td>4</td><td>5</td><td colspan="2">Characters 6 7</td><td>8</td><td>9</td><td>10</td><td>11</td><td>12</td><td>13</td></tr><tr><th><i>Ivodea</i></th><td>0</td><td>0, 1, 2</td><td>1, 2</td><td>0, 1</td><td>1</td><td>1</td><td>2</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Leionema</i> A</th><td>0</td><td>0</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0, 1</td><td>0</td></tr><tr><th><i>Leionema</i> B</th><td>0</td><td>0</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0, 1</td><td>0</td></tr><tr><th><i>Limonia</i></th><td>1</td><td>0</td><td>1, 2, 3</td><td>1</td><td>0</td><td>4</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Lunasia</i></th><td>0</td><td>0</td><td>0</td><td>0</td><td>1</td><td>0</td><td>2</td><td>0, 1</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Luvunga</i></th><td>0, 1</td><td>0</td><td>0, 1, 2</td><td>1</td><td>0</td><td>0, 1</td><td>0</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Maclurodendron</i></th><td>0</td><td>1</td><td>1</td><td>1</td><td>0</td><td>1</td><td>1</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Macrostylis</i></th><td>0</td><td>0, 1</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Medicosma</i></th><td>0, 1</td><td>0, 1, 2</td><td>1</td><td>0, 1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Melicope</i> A</th><td>0, 1</td><td>1</td><td>1</td><td>0, 1</td><td>1</td><td>1</td><td>3</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Melicope</i> B</th><td>0, 1</td><td>1</td><td>1</td><td>0</td><td>0, 1</td><td>1</td><td>3</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Melicope</i> C</th><td>0, 1</td><td>1, 2</td><td>1</td><td>0, 1</td><td>0, 1</td><td>1</td><td>3</td><td>2</td><td>0, 1, 2</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Melicope</i> D</th><td>0, 1</td><td>1</td><td>1</td><td>0</td><td>1</td><td>0, 1</td><td>3</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Melicope</i> E (<i>Platydesma</i>)</th><td>0</td><td>0, 1</td><td>1</td><td>1</td><td>0</td><td>2, 3</td><td>3</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Merope</i></th><td>0</td><td>0</td><td>2</td><td>1</td><td>0</td><td>1, 2</td><td>0</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Merrillia</i></th><td>1</td><td>0</td><td>2</td><td>1</td><td>0</td><td>3</td><td>0</td><td>0</td><td>?</td><td>0</td><td>1</td><td>0</td><td>0</td></tr><tr><th><i>Metrodorea</i></th><td>0, 1</td><td>1</td><td>2</td><td>0</td><td>0</td><td>1</td><td>2</td><td>0, 1</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Microcitrus</i> (<i>Citrus</i> s.l.)</th><td>0</td><td>0</td><td>1, 2, 3</td><td>2</td><td>0</td><td>1, 2, 3</td><td>5</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Microcybe</i></th><td>0</td><td>0</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Micromelum</i></th><td>0, 1</td><td>0</td><td>2</td><td>1</td><td>0</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Monanthocitrus</i></th><td>0</td><td>0</td><td>2</td><td>1</td><td>0</td><td>1, 2, 3</td><td>0</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Muiriantha</i></th><td>0</td><td>0</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>1</td><td>0</td><td>0</td><td>1</td><td>0</td></tr><tr><th><i>Murraya</i></th><td>1</td><td>0</td><td>1, 2</td><td>1</td><td>0</td><td>0, 1</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Myrtopsis</i></th><td>0</td><td>1</td><td>1, 2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>?</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Naringi</i></th><td>0, 1</td><td>0</td><td>1, 2</td><td>1</td><td>0</td><td>0, 1</td><td>5</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Nematolepis</i></th><td>0</td><td>0</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0, 1</td><td>0</td></tr><tr><th><i>Neobyrnesia</i></th><td>0</td><td>1</td><td>1</td><td>0</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Neoraputia</i></th><td>0, 1</td><td>0, 1</td><td>2</td><td>0</td><td>0</td><td>1</td><td>2</td><td>1</td><td>?</td><td>0</td><td>1</td><td>1</td><td>0</td></tr><tr><th><i>Neoschmidea</i></th><td>0</td><td>0</td><td>1, 2</td><td>1</td><td>1</td><td>1</td><td>2</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Orixa</i></th><td>0</td><td>0</td><td>1</td><td>0</td><td>1</td><td>0</td><td>2</td><td>1</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Pamburus</i></th><td>0</td><td>0</td><td>1, 2</td><td>1</td><td>0</td><td>1</td><td>0</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Paramignya</i></th><td>0</td><td>0</td><td>1, 2</td><td>1</td><td>0</td><td>0, 1</td><td>0</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Peltostigma</i></th><td>0, 1</td><td>0</td><td>0, 1, 2, 3</td><td>1, 2</td><td>0</td><td>0, 1</td><td>2</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Pentaceras</i></th><td>1</td><td>0</td><td>2</td><td>1</td><td>1</td><td>1</td><td>1, 4</td><td>1</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Perryodendron</i></th><td>0</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>3</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Phebalium</i></th><td>0</td><td>0</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>1, 2</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Phellodendron</i></th><td>1</td><td>1</td><td>2</td><td>0</td><td>0</td><td>0</td><td>1</td><td>1</td><td>2</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Philotheca</i> A</th><td>0</td><td>0</td><td>1, 2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>1, 2</td><td>0</td><td>0</td><td>0, 1</td><td>0</td></tr><tr><th><i>Philotheca</i> B</th><td>0</td><td>0</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>1, 2</td><td>0</td><td>0</td><td>0, 1</td><td>0</td></tr><tr><th><i>Phyllosma</i></th><td>0</td><td>0</td><td>2</td><td>0, 1</td><td>1</td><td>0, 1</td><td>2</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Picrella</i></th><td>0, 1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>0, 1</td><td>1</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Pilocarpus</i></th><td>0, 1</td><td>0</td><td>1, 2</td><td>0</td><td>0</td><td>0, 1</td><td>2</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Pitavia</i></th><td>0</td><td>1, 2</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Pitaviaster</i></th><td>0</td><td>1</td><td>1</td><td>0</td><td>1</td><td>1</td><td>1</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th>Taxon</th><td>1</td><td>2</td><td>3</td><td>4</td><td>5</td><td colspan="2">Characters 6 7</td><td>8</td><td>9</td><td>10</td><td>11</td><td>12</td><td>13</td></tr><tr><th><i>Pleiospermium</i></th><td>0, 1</td><td>0</td><td>1, 2</td><td>1</td><td>0</td><td>1</td><td>5</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Plethadenia</i></th><td>1</td><td>0, 1</td><td>1</td><td>1</td><td>2</td><td>0</td><td>2</td><td>?</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Poncirus</i> (<i>Citrus</i> s.l.)</th><td>1</td><td>0</td><td>1, 2, 3</td><td>2</td><td>0</td><td>1, 2, 3</td><td>5</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Psilopeganum</i></th><td>1</td><td>0</td><td>1, 2</td><td>1</td><td>1</td><td>2, 3</td><td>2</td><td>2</td><td>?</td><td>2</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Ptaeroxylon</i></th><td>1</td><td>1</td><td>1</td><td>0</td><td>0</td><td>0</td><td>2</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>1</td></tr><tr><th><i>Ptelea</i></th><td>1</td><td>0</td><td>1, 2, 3</td><td>0</td><td>0</td><td>1</td><td>2, 4</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Rauia</i></th><td>0, 1</td><td>0</td><td>2</td><td>0</td><td>0</td><td>1</td><td>2</td><td>1</td><td>?</td><td>0</td><td>0</td><td>1</td><td>0</td></tr><tr><th><i>Ravenia</i></th><td>0, 1</td><td>1</td><td>2</td><td>0</td><td>1</td><td>1</td><td>2</td><td>1</td><td>?</td><td>0</td><td>1</td><td>1</td><td>0</td></tr><tr><th><i>Rhadinothamnus</i></th><td>0</td><td>0</td><td>2</td><td>1</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0, 1</td><td>0</td></tr><tr><th><i>Ruta</i></th><td>1</td><td>0</td><td>1, 2</td><td>1</td><td>0</td><td>4</td><td>2</td><td>2</td><td>0, 1, 2</td><td>1, 2</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Sarcomelicope</i></th><td>0</td><td>1, 2</td><td>1</td><td>1</td><td>0, 1</td><td>1</td><td>1</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Severinia</i></th><td>0</td><td>0</td><td>1, 2, 3</td><td>1</td><td>0</td><td>0, 1</td><td>5</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Sheilanthera</i></th><td>0</td><td>0</td><td>2</td><td>0</td><td>1</td><td>0, 1</td><td>2</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Sigmatanthus</i></th><td>1</td><td>0</td><td>2</td><td>0</td><td>1</td><td>1</td><td>2</td><td>1</td><td>?</td><td>0</td><td>1</td><td>1</td><td>0</td></tr><tr><th><i>Skimmia</i></th><td>0</td><td>0</td><td>1, 2, 3</td><td>0</td><td>0</td><td>0</td><td>1</td><td>2</td><td>1, 2</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Sohnreyia</i></th><td>1</td><td>0</td><td>2</td><td>0</td><td>0</td><td>0, 1</td><td>4</td><td>0, 1, 2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Spathelia</i></th><td>1</td><td>0</td><td>2</td><td>0</td><td>0</td><td>0, 1</td><td>4</td><td>0, 1, 2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Spiranthera</i></th><td>1</td><td>0</td><td>2</td><td>0</td><td>0</td><td>1</td><td>2</td><td>1</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Swinglea</i></th><td>1</td><td>0</td><td>2</td><td>1</td><td>0</td><td>4</td><td>0</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Tetractomia</i></th><td>0</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>3</td><td>2</td><td>?</td><td>0</td><td>0</td><td>0</td><td>1</td></tr><tr><th><i>Tetradium</i></th><td>1</td><td>1</td><td>1, 2</td><td>0</td><td>1</td><td>0, 1</td><td>3</td><td>2</td><td>1, 2</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Thamnosma</i></th><td>0</td><td>0</td><td>1</td><td>1</td><td>0</td><td>2, 3</td><td>2</td><td>2</td><td>0</td><td>0, 1</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Toxosiphon</i></th><td>0, 1</td><td>0</td><td>2</td><td>0</td><td>0</td><td>1</td><td>2</td><td>1</td><td>?</td><td>0</td><td>1</td><td>1</td><td>0</td></tr><tr><th><i>Triphasia</i></th><td>0, 1</td><td>0</td><td>0, 2</td><td>1</td><td>0</td><td>0, 1</td><td>0</td><td>0</td><td>0, 1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Vepris</i></th><td>0, 1</td><td>0, 1</td><td>1</td><td>0, 1</td><td>0, 3</td><td>0, 1</td><td>1</td><td>2</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Wenzelia</i></th><td>0</td><td>0</td><td>2</td><td>1</td><td>0</td><td>3</td><td>0</td><td>0</td><td>?</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Zanthoxylum</i> A</th><td>0, 1</td><td>0</td><td>1, 2</td><td>0</td><td>1</td><td>1</td><td>3</td><td>1, 2</td><td>1, 2, 3</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Zanthoxylum</i> B</th><td>0, 1</td><td>0</td><td>1, 2</td><td>0</td><td>1</td><td>1</td><td>3</td><td>1, 2</td><td>1, 2, 3</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Zanthoxylum</i> C</th><td>0, 1</td><td>0</td><td>0, 1, 2, 3</td><td>0</td><td>1</td><td>1</td><td>3</td><td>1, 2</td><td>1, 2, 3</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Zanthoxylum</i> D (<i>Toddalia</i>)</th><td>0, 1</td><td>0</td><td>1, 2, 3</td><td>0</td><td>0</td><td>1</td><td>1</td><td>2</td><td>1, 2</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><th><i>Zieria</i></th><td>0, 1</td><td>1</td><td>1</td><td>0</td><td>1</td><td>0, 1</td><td>2</td><td>2</td><td>1, 2</td><td>0</td><td>0</td><td>0</td><td>0</td></tr></tbody></table><p><b>1.</b> Leaf type: (0) simple/unifoliolate; (1) compound. <b>2.</b> Phyllotaxis: (0) alternate; (1) opposite; (2) whorled. <b>3.</b> Flower merosity: (0) 3-merous; (1) 4-merous; (2) 5-merous; (3) polymerous. <b>4.</b> Stamen whorls: (0) haplostemonous; (1) diplostemonous; (2) more than two whorls. <b>5.</b> Carpel connation: (0) syncarpous; (1) syncarpous (style only); (2) apocarpous (no joined style); (3) one carpel. <b>6.</b> Ovules per locule: (0) 1; (1) 2; (2) 3–5; (3) 6–10; (4)>10. <b>7.</b> Fruit type: (0) berry; (1) drupe; (2) dehiscent with seeds detached; (3) dehiscent with seeds attached; (4) samara; (5) hesperidium. <b>8.</b> Endosperm: (0) lacking; (1) scanty; (2) copious. <b>9.</b> Chromosomes: (0) 9–10; (1) 14–18; (2) 28–36; (3) ≥64. <b>10.</b> Growth form: (0) shrubs or trees; (1) subshrubs; (2) perennial herbs. <b>11.</b> Flower symmetry: (0) actinomorphic; (1) zygomorphic. <b>12.</b> Tubular corolla: (0) petals free and not forming a tube; (1) petals coherent or connate and forming a corolla tube. <b>13.</b> Seeds: (0) not winged; (1) winged</p><p>(Continues)</p><p>(Continues)</p>
Figure 3 in A review of molecular genetic markers and analytical approaches that have been used for delimiting marine mammal subspecies and species
Figure 3. Published values of percent divergence between cetacean subspecies (black bars), species (white bars), and taxa of uncertain taxonomic status (gray bars). Values are based on mtDNA control region sequence data. Not all values represent net sequence divergence. See Table 1 for list of papers corresponding to each value. Since completing this work, Sousa species have been supported ((Mendez et al. 2013) and Inia subspecies changed.
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Allen Brain Atlas
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Annotated Behaviour and Observability Dataset (ABODe)
ABODe is a University of Edinburgh DataShare dataset for behavior classification in group-housed mice using home-cage video, identities, bounding boxes, ground-plate positions, and annotator labels.
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