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. 2011 May 3;30(11):2190–2204. doi: 10.1038/emboj.2011.126

Figure 4.

Figure 4

Sox2 promotes histone acetylation in Eed-deficient ES cells. Eed4 cKO ES cells transfected with either control or Flag-Sox2 expression vectors were cultured in the presence or absence of Tet for 4 days. (A, B) Global amounts of H3Ac, H4Ac and H3K56Ac are maintained in Sox2-expressing, Eed-deficient ES cells. Control or Sox2-expressing, Eed-deficient ES cells were subjected to immunostaining (A) or western blot analysis (B) using anti-H3K27me3, H3Ac, H4Ac and H3K56Ac antibodies. Bars, 50 μm. (C) Sox2 cannot restore H3K27me3 in the promoter regions of differentiation-associated genes in Eed-deficient ES cells. After culturing with or without Tet, the indicated cells were subjected to ChIP assay using an anti-H3K27me3 antibody, followed by qPCR using primers for the promoter region of the indicated genes. Asterisk, significant difference from Eed4 cKO cells cultured without Tet (P<0.05). (D) The relative amounts of H3Ac and H4Ac were quantitatively determined by ELISA. (E–G) Sox2 restores histone acetylation in the promoter regions of self-renewal genes in Eed-deficient ES cells. The indicated cells were subjected to ChIP assay using anti-H3Ac (E), H4Ac (F) and H3K56Ac (G) antibodies, followed by qPCR using primers for the promoter regions of the indicated genes. Asterisk, significant difference from Eed4 cKO cells cultured without Tet (P<0.05). Hash, significant difference from Eed4 cKO cells cultured with Tet (P<0.05). In all experiments, error bars indicate s.d. (n=3).