Supproting Materials for "Cross-Linked Selenoctanoic Acid Nanoplatform Enables Bidirectional Regulation of Intra- and Extracellular ROS: A New Avenue for Enhanced Cancer Immunotherapy" - section B

<h1><strong>Table of Content</strong></h1> <h1>1.&nbsp;&nbsp;&nbsp;&nbsp; Synthesis and characterization</h1> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.1.&nbsp;&nbsp;&nbsp;&nbsp; SeLA</h2> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.1.0.&nbsp; &nbsp; &nbsp;Lab book pages</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.1.1.&nbsp;&nbsp;&nbsp;&nbsp; NMR</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.1.2.&nbsp;&nbsp;&nbsp;&nbsp; HR-MS</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.1.3.&nbsp;&nbsp;&nbsp;&nbsp; UV</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.1.4.&nbsp;&nbsp;&nbsp;&nbsp; IR</h3> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.2.&nbsp;&nbsp;&nbsp;&nbsp; cSeLAN</h2> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.2.0.&nbsp; &nbsp; &nbsp;Lab book pages</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.2.1.&nbsp; &nbsp; &nbsp;NMR</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.2.2.&nbsp;&nbsp;&nbsp;&nbsp; UV</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.2.3.&nbsp;&nbsp;&nbsp;&nbsp; IR</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.2.4.&nbsp;&nbsp;&nbsp;&nbsp; GPC</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.2.5.&nbsp;&nbsp;&nbsp;&nbsp; MALDI-TOF-MS</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.2.6.&nbsp;&nbsp;&nbsp;&nbsp; DLS</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.2.7.&nbsp;&nbsp;&nbsp;&nbsp; Critical melle cncentration</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.2.8.&nbsp;&nbsp;&nbsp;&nbsp; ICP-MS</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.2.9.&nbsp;&nbsp;&nbsp;&nbsp; TEM</h3> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.3.&nbsp;&nbsp;&nbsp;&nbsp; cLAN</h2> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.3.0.&nbsp; &nbsp; &nbsp;Lab book pages</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.3.1.&nbsp;&nbsp;&nbsp;&nbsp; NMR</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.3.2.&nbsp;&nbsp;&nbsp;&nbsp; UV</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.3.3.&nbsp;&nbsp;&nbsp;&nbsp; DLS</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.3.4.&nbsp;&nbsp;&nbsp;&nbsp; GPC</h3> <h1>2.&nbsp;&nbsp;&nbsp;&nbsp; <em>In vitro</em> intra- and extracellular ROS regulation</h1> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.0.&nbsp; &nbsp; &nbsp;Lab book pages</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.1.&nbsp; &nbsp; &nbsp;Raman scattering spectra</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.2.&nbsp;&nbsp;&nbsp;&nbsp; HPLC</h2> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.2.1.&nbsp;&nbsp;&nbsp;&nbsp; Standard peaking</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.2.2.&nbsp;&nbsp;&nbsp;&nbsp; HPLC analysis</h3> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.3.&nbsp;&nbsp;&nbsp;&nbsp; ICP-MS</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.4.&nbsp;&nbsp;&nbsp;&nbsp; MTT</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.5.&nbsp;&nbsp;&nbsp;&nbsp; Intracellular ROS generation</h2> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.5.1.&nbsp; Fluorescence images</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.5.2.&nbsp; Flow fluorescence quantification</h3> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.6.&nbsp;&nbsp;&nbsp;&nbsp; Intracellular pro-oxidation mechanism</h2> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.6.1.&nbsp; Quantification of intracellular ATP</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.6.2.&nbsp; Quantification of intracellular NADH</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.6.3.&nbsp; Quantification of intracellular <strong>&middot;</strong>OH</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.6.4.&nbsp; Quantification of intracellular ROS</h3> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.7.&nbsp;&nbsp;&nbsp;&nbsp; Quantification of extracellular ROS</h2> <h1>3.&nbsp;&nbsp;&nbsp;&nbsp; <em>In vitro</em> ICD-inducing ability</h1> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 3.0.&nbsp;&nbsp; Lab book pages</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 3.1.&nbsp;&nbsp;&nbsp;&nbsp; ATP release</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 3.2.&nbsp;&nbsp;&nbsp;&nbsp; HMGB1</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 3.3.&nbsp;&nbsp;&nbsp;&nbsp; CRT</h2> <h1>4.&nbsp;&nbsp;&nbsp;&nbsp; T cell proliferation and activation</h1> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 4.0.&nbsp;&nbsp; Lab book pages</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 4.1.&nbsp;&nbsp;&nbsp;&nbsp; Flow cytometry analysis of the T cell proliferation</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 4.2.&nbsp;&nbsp;&nbsp;&nbsp; Quantification of IFN-&gamma; secretions</h2> <h1>5.&nbsp;&nbsp;&nbsp;&nbsp; Biosafety evaluation of cSeLAN</h1> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 5.0.&nbsp; &nbsp; &nbsp; &nbsp;Lab book pages</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 5.1.&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Hemolytic and hemagglutination assay</h2> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 5.1.1.&nbsp; Hemolytic assay</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 5.1.2.&nbsp; Hemagglutination assay</h3> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 5.2.&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Acute toxicity test</h2> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 5.2.1.&nbsp; The amounts of dead mice</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 5.2.2.&nbsp; Body weight of mice</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 5.2.3.&nbsp; Hematological assay</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 5.2.4.&nbsp; Blood biochemistry</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 5.2.5.&nbsp; H&amp;E staining of main organs</h3> <h1>6.&nbsp;&nbsp;&nbsp;&nbsp; Pharmacokinetic evaluation of cSeLAN</h1> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;6.0.&nbsp;&nbsp; Lab book pages</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;6.1&nbsp;&nbsp;&nbsp; Pharmacokinetic evaluation of cSeLAN</h2> <h1>7.&nbsp;&nbsp;&nbsp;&nbsp; <em>In vivo</em> anti-tumor immune response</h1> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;7.0.&nbsp;&nbsp; Lab book pages</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;7.1.&nbsp;&nbsp; Chemiluminescence images</h2> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 7.1.1.&nbsp; Chemiluminescence images of isoluminol</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 7.1.2.&nbsp; Chemiluminescence images of luminol</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 7.1.3.&nbsp; Related quantifications upon various treatments</h3> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;7.2.&nbsp;&nbsp; Immunofluorescence images</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;7.3.&nbsp;&nbsp; T cell proliferation</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;7.4.&nbsp;&nbsp; DC proliferation</h2> <h1>8.&nbsp;&nbsp;&nbsp;&nbsp; <em>In vivo</em> anti-tumor efficacy</h1> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;8.0.&nbsp;&nbsp; Lab book pages</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;8.1.&nbsp;&nbsp; Tumor growth</h2> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 8.1.1.&nbsp; Tumor volume</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 8.1.2.&nbsp; Tumor weight</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 8.1.3.&nbsp; Tumor images</h3> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;8.2.&nbsp;&nbsp; Body weight</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;8.3.&nbsp;&nbsp; Lung tissues with metastatic nodules</h2> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 8.3.1.&nbsp; Lung tissues images</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 8.3.2. Number of pulmonary metastatic nodules</h3> <h3>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 8.3.3.&nbsp; H&amp;E</h3> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;8.4.&nbsp;&nbsp; H&amp;E of tumor and main organs</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;8.5.&nbsp;&nbsp; Cytokine secretions</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;8.6.&nbsp;&nbsp; Kaplan&minus;Meier survival curves</h2> <h1>9.&nbsp;&nbsp;&nbsp;&nbsp; <em>In vitro</em> NK cytotoxicity and activity</h1> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;9.0.&nbsp;&nbsp; Lab book pages</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;9.1.&nbsp;&nbsp; NK cytotoxicity</h2> <h2>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;9.2.&nbsp;&nbsp; NK activity</h2>