FIGURE 5.

PD-1 is downregulated in RORγt-deficient Tregs in a T-bet–dependent manner. Naive CD4⁺ T cells sorted from Foxp3^WT, Foxp3^ΔRORγt, and Foxp3^{ΔRORγtΔT-bet} mice, were differentiated under Treg conditions and analyzed for surface expression of GITR, CD25, PD1, and both surface and intracellular expression of CTLA-4 along with Foxp3 gated on live CD4⁺ T cells at day 4 by flow cytometry, as shown in representative FACS plots (A) and scatterplots showing the percentage expression of GITR, CTLA-4, CD25, and PD-1 in gated Foxp3-expressing cells of the indicated groups (B). Data in (B) are shown as mean ± SEM (n = 6). Data are representative of two independent experiments. For p values, one-way ANOVA followed by Tukey post hoc test used in (B). *p < 0.001. n.s, not significant.