Fig. 7: Deletion of Eed promotes unrestricted fate potential in naive ESCs.

a RT-qPCR analysis for Eed and Cdx2 in Eed^fl/− (heterozygous) and Eed^−/− (null) naive ESCs. Relative mRNA expression was normalized against that of the Eed^fl/- naive ESCs, which was arbitrarily set 1. Data were presented as mean ± SEM from three biological replicates. P values were calculated using the two-sided Student’s t test. b Left, confocal images of H3K27me3 (green), CDX2 (red) and nuclei (DAPI blue) immunofluorescence in the Eed^fl/− and Eed^−/− naive ESCs. Scale bar,10 µm. Right, scatter plot showing per-nucleus intensity of CDX2 and H3K27me3 shown from the left. A total of 145 nuclei (69 nuclei for Eed^fl/- and 76 nuclei for Eed^−/−) from one representative experiment were shown. c MA plots showing up- (red) and down-regulated (blue) genes in the Eed^−/− naive ESCs relative to Eed^fl/− naive ESCs. 2C genes were selectively highlighted. RNA-seq data from biological duplicates were used to determine differential gene expression ([FDR] < 0.05). d RT-qPCR analysis showing elevated expression of MERVL and 2C genes in the Eed^−/− naive ESCs. Relative mRNA expression was normalized against that in Eed^fl/- naive ESCs, which was arbitrarily set 1. Data were presented as mean ± SEM of three biological replicates. P values were calculated by the two-sided Student’s t test. e Heatmap (Z-score) showing expression levels of trophectoderm genes in the Eed^fl/− and Eed^−/− naive ESCs. RNA-seq data from biological duplicates were shown. f GO analysis for genes significantly upregulated in the Eed^−/− naive ESCs. GO terms with fold enrichment > 2 and the [FDR] < 0.05 (defined by the Fisher’s Exact Test) were considered as significant enrichment. g Heatmap (Z-score) showing expression of genes associated with ectoderm, mesoderm and endoderm in indicated cell types. RNA-seq data from biological duplicates were shown. Source data are provided as a Source Data file.