Fig. 2

IL-1β and nitric oxide induces Foxo1 in Th9 cells. a–c Naive CD4⁺CD62L⁺CD44⁻ T cells were isolated from WT mice and then treated under Th9 (TGF-β1 + IL-4) or Th0 (without skewing cytokines) conditions with/without IL-1β or nitric oxide donor (NOC-18, 50 μM) as indicated. a After 72 h, cells were restimulated and then intracellular expression of IL-9 and IL-17 was determined by flow cytometry. b, c After 72 h, cells were harvested, RNA was prepared and qPCR was performed for determining the expression of Il9 and Foxo1. d Wt and Nos2−/− naive CD4⁺ T cells were isolated and cultured into Th9 condition, qPCR was performed for the expression of Il9 and Foxo1. e Naive CD4⁺ T cells were stimulated and polarized with TGF-β1 + IL-4 or with TGF-β3 + IL-4, Il9, Foxo1 and Irf4 mRNA was determined by qPCR. Contour plot is representative of two experiments a. Bar shows mean ± s.d. from combined three b–e experiments. *P < 0.05 **P < 0.01, ***P < 0.001 (Two-way variance analysis (analysis of variance))