Figure 2. Effects of IL-33 on iT_reg and tT_reg cells.

a, Naïve CD4⁺ T cells were cultured with anti-CD3/CD28 plus indicated cytokines and the frequencies and absolute numbers of Foxp3⁺ T cells determined 3 days later (mean ± s.e.m. of three independent experiments). b, Naïve CD4⁺ T cells were cultured for 48h with anti-CD3/CD28 plus TGF-β₁ followed by stimulation with IL-33 for 45 minutes. Blots are representative of two independent experiments. c-d, Cells were cultured and stimulated as in (b) and recruitment of GATA3 or RNA polymerase II (Pol II) to the indicated regions assessed by ChIP-qPCR. Data are from one experiment representative of two (mean ± s.d.). e, Representative plots of T_reg cells cultured with anti-CD3/CD28 plus indicated cytokines and analyzed after 3 days. Data are representative of three independent experiments. f, T_reg cells were cultured with anti-CD3/CD28 for 24h followed by stimulation with IL-33. Blots are representative of three independent experiments. g, Mixed chimaeras were generated containing WT and St2^−/− bone marrow cells. Reconstituted mice were analyzed at steady state or two weeks after infection with Helicobacter hepaticus and anti-IL-10R treatment (inflamed). Absolute numbers of WT or St2^−/− T_reg cells in steady state (n = 3) and inflamed (n = 6) hosts (mean ± s.e.m). h, Analysis of Foxp3 expression in T_reg cells from inflamed chimaeric hosts presented as geometric mean fluorescence intensity (gMFI). *P<0.05, **P<0.01 *** P<0.001 as calculated by 1way-ANOVA with Bonferroni post-test or paired Students’ t-test.