Fig. 1

Epigenetic activation of Rassf1A drives ESC differentiation via GATA1. a RASSF1A and Pou5f1/Oct4 mRNA levels in undifferentiated and differentiating human ESC lines H1 and H9. Data obtained from published GSE54186 data set. b Rassf1A mRNA levels increase upon LIF withdrawal-mediated differentiation of mouse ESC (mESC) lines V6.5 and J1. Data obtained from published GSE3749 and GDS2672 data sets. c Rassf1A mRNA (bars) and protein (blots) levels from undifferentiated (+LIF) versus differentiated (-LIF) E14Tg2a mESC. See also Supplementary Fig. 1b and Supplementary Movie 1. d Rassf1A CpG island promoter methylation (assessed by qPCR for methylated/unmethylated DNA ratio) and e H3K4me3 levels in pluripotent (+LIF) versus differentiated (-LIF) mESC. See also Supplementary Fig. 1a. f ENOCDE transcription factor ChIP data showing specific binding of GATA1 on the Rassf1A promoter of differentiated hESC and GATA1 ChIP on the Rassf1A promoter of pluripotent (+LIF) versus differentiated (-LIF) ESC. g qPCR for Rassf1A and Oct4 mRNA levels in response to siRNA-mediated Gata1 silencing in the presence or absence of LIF in ESC. Western blotting and qPCR indicate the level of Gata1 KD. See also Supplementary Figs 1d–e, 2c and 5d. h Representative ESC colonies from indicated conditions subject to LIF withdrawal assay. Scale bars: 25–50 μm. **P < 0.01 of Student’s t-test. Error bars indicate s.e.m. Data shown are representative of at least three independent experiments