Fig. 2

Impaired LEFTY-NODAL Signaling Pathway in TKO hESCs. A Gene set enrichment analysis (GSEA) highlights the similarities between TKO day4 embryoids and mouse Tet-TKO E6.5 epiblasts. Genes upregulated in Tet-TKO E6.5 epiblasts are highly biased to be upregulated in TKO day4 embryoids; B Upregulated or downregulated genes between TKO day4 embryoids and mouse Tet-TKO E6.5 epiblasts by GSEA. LEFTY1, LEFTY2 and NANOG were highlighted by red box; C RNA-seq TPM of NODAL, LEFTY1 and LEFTY2 in each cell line; D Heat map showing DNA demethylation dynamics of NODAL, LEFTY1, LEFTY2 promoters in each sample, black boxes indicate the differential methylation level, * represent compared to WT_d4 group p < 0.05; E Analysis of the percentage of 5mC at the NODAL, LEFTY1, LEFTY2 promoters by Epimark in hESCs and day 4 embryoids upon hPGCs induction; n = 3 independent experiments. Data are presented as means ± s.d. Statistical analysis was performed by Student’s t-test (two-sided): * represent compared to WT_d4 group p < 0.05. F RNA-seq counts of Nodal, Lefty1, Lefty2 in mice WT E6.25 epiblast and TKO E6.25 epiblast; G RNA-seq TPM of NANOG, POU5F1, SOX2 in hESC, WT day4 hPGCLCs and TKO day4 embryoids