Fig. 1

Naïve CD4⁺ T cells could be differentiated into Th22 cells with multiple cytokines in vitro. Naïve CD4⁺ T cells were collected from mice and cultured for 6 days in conditions designed to induce Th22 differentiation (anti-CD3/CD28 Abs, anti-IFN-γ Ab, anti-IL-4 Ab, IL-6, TNF-α, and IL-23). a Representative plots of naïve CD4⁺T cells stimulated for 6 days under optimal Th22 conditions. CD4⁺ T cells and Th (CD4⁺ IFN-γ⁻) were gated by flow cytometry to analyze the Th22 cells. b Percentage quantitation of Th22 cells. c The alterations of Nicd, Hes-1, Ahr, and Il-22 mRNAs were evaluated by RT-PCR. d Western blotting of the expression of p-STAT3, STAT3, NICD, HES-1, AHR, and IL-22 in total protein lysates from different treatment cells. e–i Representative densitometric quantification of p-STAT3, STAT3, NICD, HES-1, AHR, and IL-22 expression in T cells, β-ACTIN was used as an endogenous control for protein expression. The results show a typical experiment; each bar represents the mean ± S.E.M. of at least three independent experiments. **p < 0.01, ***p < 0.001, compared with control group