Fig. 2

Overexpression of Id2 in vitro fails to stabilize iT_reg lineage commitment, and convert them into T_H17-like cells. a Naive CD4⁺ T cells were sorted from wild-type C57BL/6 (B6) mice, and transduced with control vector (Empty RV) or vector encoding Id2 cDNA (Id2 RV) under iT_reg differentiation condition. After 3 days cells were harvested and Id2 expression was measured by RT-qPCR (n = 3, per group) and Western blot analysis. b Comparison of mRNA expression for T-bet, Gata3, Rorγt and Foxp3 between Empty RV and Id2 RV transduced T cells after 3 days post spinfection under iT_reg differentiation condition (n = 3, per group). c Flow cytometry analysis of Foxp3 expression between Empty RV or Id2 RV transduced (GFP⁺) and non-transduced (GFP⁻) iT_reg cells (n = 3, per group). d Comparison of mRNA expression for genes encoding the indicated cytokines between Empty RV and Id2 RV transduced T cells after 3 days post spinfection under iT_reg differentiation condition (n = 3, per group). e Flow cytometry analysis of IL-17A, IL-17F and IL-22 from Empty RV or Id2 RV transduced CD4⁺GFP⁺ iT_reg cells (n = 3, per group). NS, not significant, *P < 0.05, **P < 0.005 (Student’s t-test). All data are representative two or three independent experiments (error bars, s.d.)