Fig. 1
Epigenetic regulation of GDF6 during H1 hESC differentiation towards MSCs. a Gene ontology enrichment analysis of genes associated with EZH2 within 3 kb of their TSS as indicated with the GREAT annotation tool. The GO terms include PANTHER pathways. Bars represent −log10 of binomial raw P values. b Heatmap representing the expression of the indicated TGF-β signaling genes enriched for EZH2. c GDF6 mRNA was highly expressed in MSCs compared with that in hESCs. d qRT-PCR analysis of GDF6 mRNA levels in H1 hESCs treated with DMSO or 10 μmol·L−1 GSK126. e Genome browser views of EZH2, H3K4me3, and H3K27me3 peaks along the GDF6 gene. For EZH2, enrichment data were only available in H1 hESCs. For histone markers, enrichment data are presented in H1 hESCs and H1 hESC-derived MSCs. f EZH2 ChIP-qPCR for GDF6 in siScramble- and siEZH2-transfected H1 hESCs. g H3K27me3 ChIP-qPCR for GDF6 in siScramble- and siEZH2-transfected H1 hESCs. h H3K27me3 ChIP-qPCR for GDF6 in control and GSK126-treated H1 hESCs. i EZH2 ChIP-qPCR for GDF6 in control and GSK126-treated H1 hESCs. All in vitro experiments were performed independently three times. The data are expressed as the means ± SDs. *P < 0.05, **P < 0.01, and ***P < 0.001. For d, Student’s t test; for f–i, two-way ANOVA with Tukey’s post hoc test
