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. Author manuscript; available in PMC: 2020 Feb 1.
Published in final edited form as: Cancer Immunol Res. 2019 Jun 25:10.1158/2326-6066.CIR-18-0711. doi: 10.1158/2326-6066.CIR-18-0711

Fig. 5: IRF1 deficient tumor cells affect infiltrating T cell functions ex vivo and are more vulnerable to CD8+ T-cell cytotoxicity in vitro.

Fig. 5:

(A) Intracellular cytokine production of infiltrating CD8+ T cells in B16-F10 WT and IRF1-KO tumors (n=9, 8). Single cell suspension was prepared after tumor collection. The mixture of tumor cells and infiltrated lymphocytes was stimulated overnight with PMA and ionomycin, then cytokine secretion was blocked with GolgiPlug™ for 4 hrs. Representative flow-cytogram for intracellular cytokine expression is shown. For each type of cytokine expression, cumulative mean and SEM were plotted from twice repeated experiments, statistical significance was calculated by unpaired student t test, and presented as * P < 0.05. (B) The percentage of dead tumor cells at different Pmel T : tumor cell ratios. 10,000 tumor cells in each 96-well plate were cocultured with different amount of Pmel CD8+ T cells overnight. Cells were harvested and examined for viability using Zombie Aqua Fixable Viability Kit. The experiment was repeated 3 times. For each data point mean and SEM from twice repeated experiments were plotted, statistical significance calculated by two-way ANOVA with Sidak’s multiple comparison test, and presented as * P < 0.05, ** P < 0.01.