Figure 4.

p300 is recruited to the DUX4 binding site in ZSCAN4. (A) Location of PCR amplicons in the ZSCAN4 gene used for ChIP analyses. The previously-identified DUX4-binding site is at +1430, in exon 2. (B–G) ChIP assays with various antibodies at the indicated ZSCAN4 loci (ZSCAN4-P: promoter region, ZSCAN4-E: exon 2 region). LHCN-M2 cells were treated with 250 ng/ml dox for 6 h to express DUX4 or DUX4-ΔC and subjected to ChIP analyses.% input of H3K18Ac, H3K27Ac, and H3K4me3 were normalized with% input of H3 (n = 3, error bars represent SEM). Note that both DUX4 and DUX4ΔC are recruited to the DUX4 site in exon 2, but p300 is not recruited by DUX4ΔC, and acetylation of H3K18 is not increased by DUX4ΔC.