Figure 7: Bach2 inhibits the formation of a positive feedback loop to induce Th2 cell development.

(a) Global patterns of Bach2 binding to the Batf gene locus in the wild-type (WT) and Bach2-deficient naive cells cultured under neutral conditions for 5 days were determined using a ChIP-seq analysis. (b) Results of the ChIP assay with a quantitative PCR (qPCR) analysis of Bach2 binding to the Batf gene locus in the WT naive CD4 T cells cultured under IL-2 conditions for 48 h; the results are presented relative to those of input DNA with the s.d. **P<0.01 (Student’s t-test; n=3). (c) Results of the ChIP assay with a qPCR analysis of Batf or Irf4 binding at the Batf #1 and Batf #2 regions in the WT and Bach2-deficient CD4 T cells cultured under neutral conditions for 48 h; the results are presented relative to those of input DNA with the s.d. *P<0.05 and **P<0.01 (Student’s t-test; n=3). The data are representative of three-independent experiments with similar results. (d) The levels of histone H3K4 tri-methylation at the transcription start sites of the Batf and Batf3 gene loci in the WT or Bach2-deficient naive CD4 T cells cultured under IL-2 conditions for 48 h 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=3). (e) Results of the quantitative reverse transcription (RT)–PCR analysis of Batf and Batf3 mRNA in the WT, Bach2-, Stat6- or Bach2/Stat6 double-deficient naive CD4 T cells stimulated with an anti-TCR-β mAb plus an anti-CD28 mAb in the presence of IL-2 for the indicated hours. The results are presented relative to the mRNA expression of Cd3ɛ, with the s.d. **P<0.01 (Student’s t-test; n=3). (f) Results of the quantitative RT–PCR analysis of Bach2 and Batf3 mRNA in the WT or Bach2 TG naive CD4 T cells cultured under Th2 conditions for 48 h. The results are presented relative to the mRNA expression of Cd3ɛ, with the s.d. **P<0.01 (Student’s t-test; n=3).