Fig. 5: Cooperation between STAT5 and BATF in the plasticity of the Il9 locus.

Naive CD4+ T cells were isolated from the spleen and differentiated into Th lineage for 5 days. Retrovirus expressing caSTAT5 (tagged with hNGFR), BATF (tagged with Thy1.1), or IRF4 (tagged with GFP) was transduced on day 1. For cytokine production analysis, cells were stimulated with PMA/Ionomycin for 5 h and monensin was added for the last 2 h. a Venn diagram of overlap between BATF and STAT5b target genes in Th9 cells. b, c Flow cytometric analysis of cytokine expression in Th17 cells co-transduced with caSTAT5 and BATF retrovirus. d Flow cytometric analysis of IL-9 expression in Th17 cells co-transduced with caSTAT5 and IRF4 retrovirus; cells were analyzed on day 5. e, f Flow cytometric analysis of cytokine expression in WT or Batf^ΔZ/ΔZ Th9 cells co-transduced with caSTAT5 and BATF retrovirus; cells were analyzed on day 5. g, h Flow cytometric analysis of cytokine expression in WT or Batf^ΔZ/ΔZ Th17 cells co-transduced with caSTAT5 and BATF retrovirus; cells were analyzed on day 5. i Flow cytometric analysis of IL-9 expression in caSTAT5 lone or caSTAT5 and BATF retrovirus co-transduced Th0, Th1, Th2, and Treg cells, cells were analyzed on day 5. Date are mean ± SEM of three mice per experiment and representative of two independent experiments. Two-way ANOVA with Sidak’s multiple comparison test was used to generate p-values. ns: no significant, p > 0.05. See also Supplementary Fig. 4.