Fig. 4.

RORγt antagonizes cytotoxic functions in CD8⁺ T cells. (A and D) RT-qPCR experiments assess expression of Gzmb and Gzmk (A) or T-bet and Eomes (D) from WT (black bars) or Stat3^−/− (gray bars) cells cocultured in Tc17 conditions as described in Fig. S2A, transduced with RORγt or control (empty) retroviruses, and sorted for CD45 allele expression before RNA preparation. Data are expressed relative to expression in WT Tc17 cells transduced with control virus (set to 1) and is representative of two mice per genotype analyzed in two independent experiments. (B) Contour plots show intracellular expression of IL-17 vs. granzyme B on WT or Stat3^−/− cells cocultured in Tc17 conditions as described in Fig. S2A and retrovirally transduced as indicated. Data are gated on transduced cells and representative of three mice per genotype in three independent experiments. (C) Overlaid histograms show intracellular granzyme B and IFNγ expression in WT (gray shaded) or Stat3^−/− (transduced as indicated) CD8⁺ T cells assessed as in B. Data are representative of three mice per genotype in three independent experiments. (E and F) WT CD8⁺ T cells were transduced with RORγt or control Thy1.1-expressing retrovirus and cultured in Tc1 conditions. (E) Contour plots show intracellular expression of IL-17 vs. granzyme B, gated on retrovirus-expressing (Thy1.1⁺) cells. (F) Before–after plots compare intracellular granzyme B and IFNγ expression in Thy1.1⁺ (empty squares) and Thy1.1^– (filled circles) cells within the same culture. Data [mean fluorescent intensity (MFI)] is expressed relative to that in Thy1.1^– cells in control virus-transduced cultures, set to 100 within each mouse. Each pair of symbols represents a separate culture; data are from five mice analyzed in three independent experiments *P < 0.05; ns, not significant.