Images used for protein quantification in Hayes et al (2019) Current Biology: Figure 3

<p>Images used for quantification of PIF4-HA and PIF5-HA protein stability in Fig 3. Hayes et. al 2018.</p> <p><em>35S:PIF4-HA </em>and <em>35S:PIF5-HA</em> (L<em>er</em>) were germinated on &frac12; MS plates in white light (16:8h photoperiod) for 3 days, before being transferred to plates with or without 75mM NaCl. Plates were then grown in red light or a further 2 days. On day 5, at Zt 3, half the plates were moved to red+ far-red light. Tissues were harvested at Zt 4. 10 seedlings were homogenized and proteins were extracted in 70 &micro;l Cracking Buffer (125mM Tris pH 7.4, 2% SDS, 10% Glycerol, 6M Urea, 5% &beta;ME) and 15 &micro;l of the extract was run on a 10% polyacrylamide gel. Blots were probed with anti-GFP-HRP (1:1000). Membranes were developed with 50/50 &lsquo;pico&rsquo; and &lsquo;femto&rsquo; chemiluminescence substrate (Thermo) on a ChemiDoc (Biorad).</p> <p>&nbsp;</p> <p>Experiment was performed on 4 dates (in duplicate).</p> <p>R= red light</p> <p>FR= red +far-red light</p> <p>RN= red light +NaCl</p> <p>FRN= red +far-red light +NaCl</p> <p>&nbsp;</p> <p>Sample order is as follows:</p> <p>&nbsp;</p> <p>01-08-18:</p> <p>PIF4-HA: R1, R2, FR1, FR2, RN1, RN2, FRN1, FRN2</p> <p>PIF5-HA: R1, R2, FR1, FR2, RN1, RN2, FRN1, FRN2</p> <p>&nbsp;</p> <p>08-08-18:</p> <p>PIF4-HA: R3, R4, FR3, FR4, RN3, RN4, FRN3, FRN4</p> <p>PIF5-HA: R3, R4, FR3, FR4, RN3, RN4, FRN3, FRN4</p> <p>&nbsp;</p> <p>15-08-18:</p> <p>PIF4-HA: R5, FR5, RN5, FRN5, R6, FR6, RN6, FRN6</p> <p>PIF5-HA: R5, FR5, RN5, FRN5, R6, FR6, RN6, FRN6</p> <p>&nbsp;</p> <p>10-10-18:</p> <p>PIF4-HA: R7, FR7, RN7, FRN7, R8, FR8, RN8, FRN8</p> <p>PIF5-HA: R7, FR7, RN7, FRN7, R8, FR8, RN8, FRN8</p>