Fig. 5

Foxo1 controls IL-9 induction in TGF-β1-induced iTregs. a–c Naive CD4⁺ T cells were activated with plate bound anti-CD3 and anti-CD28 in the presence or absence of TGF-β1. a 72 h later, cells were restimulated with PMA/Ionomycin for 5–6 h and then intracellular staining for IL-9 was performed, data representative of one of two independent experiments. At 72 h RNA was prepared from above cultured cells and qPCR was performed for Foxo1 and Klf2 b, Batf, Irf4 and Spi1 c. Naive T cells were activated with anti-CD3 and anti-CD28 in the presence of TGF-β1 with or without Foxo1 inhibitor (25 nm), Foxp3 expression was determined by flow cytometry at 72 h, bars to the right shows % Foxp3 induction d, percentage CD4⁺Foxp3⁺ cell were determined by FACS. e, f, mRNA expression of Il9 and Il17 was determined by qPCR from above experiments. Bars show mean ± s.d. from combined three b–e experiments. *P < 0.05, **P < 0.01, ***P < 0.001 (Student’s t-test)