Figure 4: Epigenetic regulation of Cxxc1 target genes in DP thymocytes.

(a) Genome browser views of murine Cd8a–Cd8b, Zap70, Lck and Rorc loci, with Cxxc1-binding signals (red) in WT DP thymocytes and H3K4me3 signals (blue) in WT and Cxxc1-deficient DP thymocytes. (b–f) Detection of Cxxc1 binding (b), Setd1 binding (c) and H3K4me3 (d), H3K9ac (e) and H3K27ac (f) modifications by ChIP–qPCR on Cd8a–Cd8b, Zap70, Lck and Rorc loci in WT and Cxxc1-deficient DP thymocytes. An intergenic region of chromatin 2 served as control. Data comes from three independently sorted sample sets (for H3K4me3, H3K9ac and H3K27ac, each from two mice; and for Cxxc1 and Setd1, each from four mice). The statistical significance was determined by unpaired t-test (two tailed). NS, not significant. **P<0.01, ***P<0.001. Error bars indicate s.d. (g) mRNA expression of Cd8a, Cd8b, Zap70, Lck and Rorc in WT and Cxxc1-deficient DP thymocytes, detected by quantitative real-time PCR. (h) Protein levels of CD8α, CD8β, ZAP70 and RORγt detected by fluorescence-activated cell sorting in control and Cxxc1-deficient DP thymocytes. Data are from three-separate mice experiments (g,h). The statistical significance was determined by unpaired t-test (two tailed). ***P<0.001 Error bars indicate s.d.