Figure 3: Decreased Th2 cell differentiation and Th2 cytokine production in Bach2-TG naive CD4 T cells.

(a) The results of the intracellular flow cytometry analysis of IL-4/IFN-γ in the wild-type (WT) and Bach2-TG naive CD4 T cells cultured under Th2 conditions. The numbers of cells are indicated in each quadrant. The data are representative of at least three-independent experiments with similar results. (b) The results of the ELISA for cytokines in the supernatants of the cells in (a) stimulated with an immobilized anti-TCR-β mAb for 16 h. **P<0.01 (Student’s t-test; mean±s.d., n=3). (c) The results of the flow cytometry analysis of Gata3 in WT or Bach2-deficient naive CD4 T cells cultured under Th2 conditions for 2 days. The data are representative of at least three-independent experiments with similar results. (d) The results of the ChIP assay of the binding of Gata3 to the RHS6 within the LCR of the Th2 cytokine gene locus, the Il4 IE and the CGRE in naive CD4 T cells cultured under Th2 conditions for 2 days; the results are presented relative to those of input DNA with the s.d. **P<0.01 (Student’s t-test; n=3). (e,f) The levels of histone H3K4 tri-methylation (e) and H3K27 acetylation (f) at the Th2 cytokine gene loci in naive CD4 T cells cultured under Th2 condition for 2 days were determined using a ChIP-qPCR assay; the results are presented relative to those of input DNA with the s.d. **P<0.01 (Student’s t-test; n=5).