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. 2017 May 26;45(15):8758–8772. doi: 10.1093/nar/gkx475

Figure 6.

Figure 6.

Chd2 is associated with Oct3/4 for H3.3 deposition in developmental gene loci. (A) Immunoprecipitations were performed from WT ESC extracts using a Chd2-specific antibody or IgG as a control. Western blot analyses of the products using antibodies against Chd2, Oct3/4, Sox2 and H3.3 are shown. (B) Motif analysis of the Chd2-binding region around the bivalent genes (±50 kbp) using HOMER (43). (C) ChIP-qPCR assay using anti-Chd2 antibodies at bivalent genes (Gata6, T, Nes, Nanog and Myh1) in WT and Oct3/4-depleted mESCs. Recovery efficiency (mean ± standard deviation of three independent experiments) is expressed as enrichment relative to the input. *P < 0.05. (D) Box plots show H3K27me3 and (E) H3.3 ChIP-seq signal intensities for bivalent and active state genes at the TSS ± 5 kbp in ZHBTc4 cells with or without doxycycline treatment. The Wilcoxon test was used for statistical analysis. (F). ChIP-qPCR assay using anti-H3K27me3 and (G) anti-H3K27ac antibodies. Both bivalent genes (Gata6, T and Nes) and an active gene (Nanog) were analyzed in WT and Oct3/4-depleted ESCs. Recovery efficiency (mean ± standard deviation of three independent experiments) is expressed as enrichment relative to the input. *P < 0.05.