Fig. 2

2C genes including Zscan4 are repressed in primed mEpiSCs. A Heatmap highlighting gene expression profile related to 2C genes in CBA × C57 P5 and P15 naïve and primed cells. The representative genes are listed on the right. B Gene set enrichment analysis (GSEA) indicating that downregulated genes under primed state are highly enriched in the 2C gene set. C Venn diagram showing the overlap between upregulated genes in naïve and primed cells and 2C genes. P value was calculated by Fisher’s exact test. D Protein level of Zscan4 by western blot analysis of two different naïve and primed cell lines at P5 and P15. MEF cells served as a negative control. E Representative images of immunofluorescence of Zscan4 protein in CBA × C57 mESCs and mEpiSCs cultures at P5 and P15. Scale bar = 10 μm. Right panel, proportion of Zscan4⁺ cells based on immunofluorescence images. Number of cells counted is shown on the top of the bar. F Gene set enrichment analysis (GSEA) indicating that downregulated genes under primed state are highly enriched in the metabolic glycolysis pathway as well as in the metabolic oxidative phosphorylation pathway. G Oxygen consumption rate (OCR) in response to oligomycin, FCCP, and rotenone/antimycin of naïve and primed cells (n = 3)