Differentiation of Ezh2-null naïve CD4⁺ T cells. (A) Schema of naïve CD4⁺ T-cell activation and differentiation conditions. (B) Four-day proliferation of naïve CD4⁺ T cells purified from Ezh2^fl/fl and Ezh2^fl/fl.CD4Cre mice that were cocultured with WT (Ly5.1⁺) naïve CD4⁺ T cells. iTreg-1 and -5 indicate the addition of 1 and 5 ng/mL TGF-β1 under iTreg conditions. Cytokine production in Th0 (D), Th2 (E), Th17 (E), and Th1 and iTreg (F) cells at day 4. (G) T-bet and Gata-3 mean fluorescent intensity (MFI) in Th0, Th1, and Th2 cells. Data are representative of 3 independent experiments for (A-G). (H) mRNA expression of Tbx21, Ifng, and Foxp3 in naïve and differentiated Th0 and iTreg cells; Rps9 was used as a reference gene. Data are presented as mean ± standard error of the mean (SEM) of 2 independent experiments.