Fig. 6
From: JunB plays a crucial role in development of regulatory T cells by promoting IL-2 signaling
CD4+ T cells lacking Junb exhibit an impairment in production of IL-2 and expression of CD25 under Treg-inducing conditions in vitro. a Naive CD4+ T cells isolated from control or Junbfl/flCd4-Cre mice were cultured for 3 days under Treg-inducing conditions in the absence or presence of recombinant IL-2 (50 ng/ml), and then analyzed by flow cytometry. b Naive CD4+ T cells isolated from the indicated mice were cultured for 9 h under the Treg-inducing conditions in the absence of exogenous IL-2. The concentrations of IL-2 in the culture supernatant were measured by ELISA. Results are means ± SDs of triplicate samples and are representative of three independent experiments. c Naive CD4+ T cells from the indicated mice were cultured for 3 days under Treg-inducing conditions in the presence of neutralizing anti-IL-2 antibody (20 μg/ml), and then analyzed by flow cytometry. d Mean fluorescence intensities (MFIs) of CD25 are shown. Pooled results of 4 or 6 independent experiments performed under the conditions in (a, c) are presented. e Naive CD4+ T cells from the indicated mice were cultured for 3 days under the Th0 conditions in the absence or presence of neutralizing anti-IL-2 antibody (20 μg/ml), and then analyzed by flow cytometry. f MFIs of CD25 are shown. Pooled results of 4 independent experiments performed under the conditions in (e) are presented. g Average percentages of Foxp3+ cells under the indicated culture conditions are presented. Results are means ± SEMs (n = 4–6 per each genotype). Representative results of flow cytometry are shown (n = 4–6 mice per each genotype) (a, c, e). h The nucleotide sequence of Il2ra promoter. Potential AP-1 binding sites are boxed. i Naive CD4+ T cells were cultured under Treg-inducing conditions for 3 days, and subjected to the ChIP assay using the indicated antibodies. The precipitated DNA was analyzed by quantitative PCR using primer pairs corresponding to the indicated genomic regions. The enrichment of the precipitated DNA relative to input is shown. Results are means ± SDs of triplicate samples and are representative of at least four independent experiments. Statistical significance was determined by two-tailed Student’s unpaired t test (b, g, i) or two-tailed Student’s paired t test (d, f). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, n.s. not significant
