Supplementary MaterialsS1 Fig: IRF4 regulates T-bet and Eomesodermin levels in activated CD8+ T cells. and the percentage of normalized MFIs for T-bet relative to Eomes. (C) Graphs display compilations of proportions and numbers of T-bet+ Eomes- and T-bet+ Eomes+ cells. Each data point represents an individual mouse and data are a compilation of three self-employed experiments; significant differences Pipequaline determined by Regular one-way ANOVA using Tukeys multiple assessment test.(TIF) pone.0144826.s002.tif (12M) GUID:?32CF7775-997B-400D-A41D-DE1B7B3CB055 S3 Fig: gene dosage regulates the proportions of virus-specific CD8+ T cells during persistent LCMV-clone 13 infection. Splenocytes from LCMV-clone 13 infected WT, and mice were harvested between D21-24 p.i. and stained having a viability dye, LCMV-specific H2-Db-GP276 and H2Db-GP33 tetramers, and Pipequaline antibodies to CD8, T-bet and Eomes. (A, C) Graphs display the figures and proportions of T-bet+ Eomes+ (remaining) and T-bet- Eomes+ (ideal) populations. Each data point represents an individual mouse and data are compilations of five self-employed experiments; significant variations determined by Regular one-way ANOVA using Tukeys multiple assessment test. (B, D) Dot plots of uninfected control and LCMV Armstrong infected control used to determine gating of T-bet versus Eomes for each tetramer stained subset.(TIF) pone.0144826.s003.tif (12M) GUID:?6EB51057-709C-4027-B6A0-852DC570029C S4 Fig: Clearance of LCMV-clone 13 leads to increased T-bet to Eomesodermin ratios. Splenocytes from LCMV-clone 13-infected WT, and mice were stained having a viability dye, LCMV-specific H2-Db-GP276 and H2Db-GP33 tetramers, and antibodies to CD8, T-bet and Eomes, and analyzed between D112-114 p.i. Graphs display the MFI of T-bet and Eomes each normalized to the average of WT samples, and the percentage of normalized MFIs for T-bet relative to Eomes, for live CD8+ H2-Db-GP276 (A) and H2-Db-GP33 (C) specific cells. Graphs display compilations of the quantities and proportions of Eomeshi PD-1hi H2-Db-GP276 (B) or Pipequaline H2-Db-GP33 (D) particular cells. Each data stage represents a person mouse and data certainly are a compilation of three unbiased experiments; significant distinctions determined by Normal one-way ANOVA using Tukeys multiple evaluation test. Icons with vivid outlines signify mice whose serum viral titers had been below the limit of recognition at D112-114 p.we.. $ denotes statistically factor between WT and samples when examining just mice with undetectable serum viral titers (vivid outlined icons). Significant distinctions between bold specified samples were dependant on unpaired t check with Pipequaline Welchs modification.(TIF) pone.0144826.s004.tif (11M) GUID:?42D3646B-432A-4353-BC3F-04DAE5FBD7C9 S5 Fig: Compound haplo-deficiency of and will not alter exhaustion marker expression, cytokine production, or effector function in H2-Db-GP276 specific cells. Splenocytes from LCMV-clone 13-contaminated WT, and mice had been stained using a viability dye, LCMV-specific H2-Db-GP276 tetramers, and antibodies to Compact disc8, T-bet, Eomes, 2B4, Compact disc160, LAG-3, PD-1, and granzyme B and examined at D22 p.we. (A) Variety of H2-Db-GP276 particular cells at D22 p.we. (B) Graphs present the proportions of 2B4-, Compact disc160-, LAG-3-, and PD-1-positive H2-Db-GP276 particular cells at D22 p.we. (C) Dot plots present T-bet versus PD-1 staining on H2-Db-GP276 particular Compact disc8+, live cells. Graph displays the proportions of T-bethi PD-1lo H2-Db-GP276 Compact disc8+ particular cells. * Indicates significant distinctions in accordance with WT examples statistically. (D) Dot plots present Eomes versus PD-1 staining on H2-Db-GP276 particular, Compact disc8+, live cells. Graph displays proportions of Eomeshi PD-1hi H2-Db-GP276 Compact EZH2 disc8+ particular cells. (E-H) Splenocytes from LCMV-clone 13-infected WT, and mice were isolated at D22 p.i. and stimulated with GP276 peptide, stained having a viability dye and antibodies to CD8, IFN, TNF and IL-2. (E) Dot plots display representative staining of WT CD8+ live cells (CD8 versus IFN) and gated IFN+ CD8+ live cells (TNF versus IL-2). (F) Graph shows the proportions of IFN+ cells gated on CD8+ live cells for each genotype. (G) Graphs display the proportions of TNF+ IL-2- (remaining) and TNF+ IL-2+ (ideal) cells gated on IFN+ CD8+ live cells for each genotype. (H) Graph shows the numbers of Granzyme B+ H2-Db-GP276 CD8+ Pipequaline live cells for each genotype. Each data point represents an individual mouse and data are compilations of three self-employed experiments; significant variations were determined by Regular one-way ANOVA using Tukeys multiple assessment test.(TIF) pone.0144826.s005.tif (39M).
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