T_reg cells from Bcl11b^F/F/CD4-Cre mice show reduced IL-10 expression and up-regulation of proinflammatory cytokines, and Bcl11b binds to the IL-10 promoter and noncoding sequence CNS+6.5. (A) IL-10 mRNA levels in T_reg cells from Bcl11b^F/F/CD4-Cre and WT mice evaluated by qRT-PCR analysis. (B) Flow cytometry analysis of IL-10, IL-17, Tnf, and Ifn-γ in T_reg cells from Bcl11b^F/F/CD4-Cre/Foxp3-GFP and Foxp3-GFP colon LP. Data are representative of four pairs of mice. (C) ChIP assays with anti-Bcl11b or IgG antibodies. The immunoprecipitated DNA was amplified with primers corresponding to the indicated regulatory regions. Binding was normalized to the input, as described in Materials and methods. Enrichment by Bcl11b is indicated as fold increase versus IgG. This is one representative experiment out of two.