Figure 4: IL-27 does not suppress RORγt expression in committed Tc17 cells.

CD8⁺ T cells of WT C57BL/6 mice were activated with anti-CD3 and anti-CD28 Abs in the presence of IL-6+TGF-β ± IL-27 and blocking Abs to IL-4 and IFN-γ (1^st stimulation). After 72 h cells were rested for 2 days in the presence of IL-2 and then restimulated during three days with Abs in the presence of the cytokines indicated in each panel (2^nd stimulation). Cells were then stained and analyzed by flow cytometry for RORγt expression (A). Real-time PCR analysis of RORγt expression in developing (B) and committed Tc17 cells (C) differentiated from naïve WT CD8⁺ T cells and reactivated in the presence of indicated cytokines. Data are representative of three (flow cytometry) and two qPCR experiments. (error bars, s.e.m.; *p< 0.05; ***p<0.001).