Journal: Biomaterials

Article Title: Biphasic response of T cell activation to substrate stiffness.

doi: 10.1016/j.biomaterials.2021.120797

Figure Lengend Snippet: Fig. 1. Characterization of the hydrogel system for T cell activation. (A) Schematic of antibody coated hydrogels as T cell activation platform. Poly acrylamide gels were polymerized with streptavidin- acrylamide, then coated with biotinylated Goat-anti- mouse and secondary activating antibodies to CD3 and CD28. Mixed CD4+/CD8+ polyclonal primary human T cells were stimulated on activating sub strates. (B) Mechanical testing via indentation indi cate hydrogels with varying ratios of acrylamide and bis-acrylamide have Young’s modulus between 5 and 110 kPa (Data are mean ± s.d., n = 4 gels for 5 kPa, n = 7 gels for all other formulations). (C) Confocal microscopy imaging of fluorescent antibodies coated on hydrogels indicate attachment of antibodies to hydrogel surface. (D) Streptavidin-acrylamide con centrations were varied to obtain similar coating of antibodies on hydrogels, as verified by fluorescence intensities on the surface of gels. (n = 3 gels for each formulation).

Article Snippet: Key resources table REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Biotinylated Goat-anti-mouse BioLegend 405303 Mouse-anti-human CD3 (OKT3) BioXCell BE0001-2 Mouse-anti-human CD28 (9.3) BioXCell BE0248 Biological Samples Leukopack New York Blood Center Chemicals, Peptides, and Recombinant Proteins Acrylamide (40%) Sigma 01697 Bis-acrylamide (2%) Fisher BP1404-250 Ammonium persulfate (APS) Sigma A3678 Tetramethylethylenediamine (TEMED) Sigma T7024 Streptavidin-acrylamide Fisher S21379 (3-Aminopropyl)triethoxysilane (APTES) Sigma 440140 Glutaraldehyde Sigma G7776 Alexa-fluor 488 Thermo A20000 Blebbistatin Sigma 203391 Prolong Diamond ThermoFisher P36961 Qiazol Qiagen 79306 Critical Commercial Assays Human IL-2 Secretion kit (APC) Miltenyi Biotec 130-090-763 RosetteSep Human T cell Enrichment Kit Stemcell Technologies 15061 CellTrace CFSE Dilution Assay ThermoFisher C34554 CellVue Jade ThermoFisher 88-0876-16 Software and Algorithms FlowJo 7.6 FlowJo LLC FCS Express V6 De Novo Software Graphpad Prism 7 Graphpad Software Kallisto Bray et al.

Techniques: Activation Assay, Confocal Microscopy, Imaging, Fluorescence, Formulation

Journal: bioRxiv

Article Title: CD8 + T cell response promotes viral clearance and reduces chances of severe testicular damage in mouse models of long-term Zika virus infection of the testes

doi: 10.1101/2024.01.22.575592

Figure Lengend Snippet: A. Histology of A129 mice testes. B. Percentage of CD3 events in blood samples of A129 mice excluding red blood cells at 2 or 18 dpi. C. Percentage of seminiferous tubules (ST) with spermatogenesis in each A129 mouse testicular sample. D-F show histology, CD3 levels, and percentage of ST for the C57BL/6J mice. Statistical significance was assessed with one-way ANOVA Sidak’s multiple comparison correction. Bars show mean and error bars represent standard deviation. Lines represent the mean. **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; n.s., no significant difference.

Article Snippet: For experiment shown in , 0.1 mg/mouse of anti-mouse CD3ε F(ab’)2 fragments (clone 145-2C11, BioXCell) or control f(ab’)2 fragments of polyclonal hamster IgG (BioXCell) were injected intraperitoneally on days -3, -1, 2, 7, and 14 relative to infection at day 0.

Techniques: Comparison, Standard Deviation