Fig. 5

a Flow cytofluorimetry analysis of TNG-A Nanog::GFP cells at 4 DIV. b GFP-positive cell ratios at 2 DIV and 4 DIV after inhibition of SmarcA4 (shSA4), DNMT (AZA), or KDM (CHQ) compared with control (Ctrl). c, d Similar analysis as in a and b, with a mESC line carrying GFP under the human Nanog promoter (HNP). e, f RT-PCR gene expression analysis. Values are relative to β-actin mRNA expression. The lowest and highest expression levels were normalized to 1 in the left and right histograms, respectively. g Nanog proximal promoter methylation. The scheme shows the region of the mouse Nanog promoter amplified for bisulfite-treated DNA sequencing (bis-seq). Black circles in grid rows indicate CpG methylation sites. The histogram shows the percentage of CpG methylation in different culture conditions as above. *p = 0.05, **p = 0.01, ***p = 0.001 (b, d, g, Student’s t-test; e, f, REST randomization test). Error bars show standard error