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. 2011 Apr 18;108(18):7391–7396. doi: 10.1073/pnas.1018279108

Fig. 5.

Fig. 5.

Effect of OCT4 overexpression on chromatin structure and architecture. A–C are the results of ChIP-qPCR analysis of CTCF, OCT4, and RAD21, respectively, at the CBS5 site using chromatin isolated from IMR90 cells overexpressing OCT4 (OCT4 OE) or a control protein (LacZ). Western blot analysis showing the efficacy of OCT4 OE in IMR90 cells is displayed in the inset of B. D shows the interaction frequency for major looping between CBS5 and the heterochromatin boundary site (p40) in IMR90 cells overexpressing OCT4 and LacZ. In E, H3K27me3 ChIP-qPCR analysis of chromatin isolated from IMR90 cells overexpressing OCT4 (OCT4 OE) was carried out at four CTCF binding sites, CBS4, 5, 6, and 7 in the HOXA locus. F is the mRNA expression analysis on the HOXA genes (HOXA6, 7, 9, and 10) after OCT4 overexpression in IMR90 cells. Values of the y axis represent the relative copy number of HOXA cDNAs normalized to ACTG1 cDNA. G is the proposed model of regulated chromatin looping at HOXA locus.