Figure 6.

Requirement for Runx complex binding to the silencer in establishment of epigenetic Thpok silencing. (A) Schematic structures of mutant Thpok^gfp reporter alleles. Black boxes, open circles marked with S, orange lines and X represent exons, the silencer region, the 242–401 core silencer sequences and mutations at Runx sites, respectively. (B) Histograms showing Thpok-GFP expression in the indicated T-cell population from mice of the indicated genotypes. Numbers in the histogram indicate the percentage of GFP-positive cells. (C, D) ChIP assays showing binding of Runx/Cbfβ complexes to the silencer region on the Thpok, the Thpok^{gfp:242–401RM} or the Thpok^gfp:3xRM allele in DP thymocytes (C), histone H3K4 tri-methylation (K4me3) and histone H3K27 tri-methylation (K27me3) to the silencer region on the Thpok or Thpok^{gfp:242–401 RM} allele in CD8⁺ T cells (D). Data are one representative of at least three individual experiments. (E) Histograms showing Thpok-GFP expression in CD8⁺ T cells of Thpok^+/gfp:3xRM mice with full (Bcl11b^+/+) or half dosage (Bcl11b^+/−) of the Bcl11b gene. Numbers in the histogram indicate the percentage of GFP-positive cells. Graph at the right shows results with individual mice.

Source data for this figure is available on the online supplementary information page.

Source Data for Fig 6C ^{(302.1KB, pdf)}

Source Data for Fig 6D ^{(268.2KB, pdf)}