Fig. 3

NOTCH signaling pathway was involved in differentiation of CD4⁺ T cells towards Th22 cells. Naïve CD4⁺ T cells were cultured in conditions designed to induce Th22 differentiation, either 1 µg/mL jagged-1 treatment alone or jagged-1 combined with DAPT (10 µM). a The percentage of Th22 (CD4⁺IL-22⁺IFNγ⁻IL-17ˉ) cells was analyzed by flow cytometry in naïve CD4⁺ T cells treated with PBS, jagged-1 or jagged-1 plus DAPT. b Percentage quantitation of Th22 cells. c Gene expression of Nicd, Hes-1, Ahr, and Ii-22 was analyzed by RT-PCR. d Representative western blot showing protein levels of p-STAT3, STAT3, NICD, HES-1, AHR, and IL-22 extracted from different groups. e−i Densitometric analysis of p-STAT3, STAT3 NICD, HES-1, AHR, and IL-22 levels was performed with Quantity One software and data were represented as the means ± S.E.M. of three different experiments. **p < 0.01, ***p < 0.001, compared with control group; ^#p < 0.5, ^##p < 0.01, ^###p < 0.001, for jagged-1 versus Th22 groups; ^△△p < 0.01, ^△△△p < 0.001, for DAPT versus jagged-1 groups