Transcriptomic and epigenetic features of bEPSCs. (A) DNA methylation levels in bEPSCs by whole-genome bisulfite sequencing analysis. Boxplots of the averaged DNA methylation levels (CpG sites) of 5 kb tiles in bEPSCiPS-Q36, bEPSCES-A6, and BFFs. The bottom and top of the boxes indicate the first and third quartiles, respectively, and the lines inside the boxes indicate the medians of the data. (B) DNA methylation in DMR of bovine-imprinted genes, including DLK1-DIO3 cluster, H19 cluster, IGF2R, MEG3, PEG3, PEG10, PLAGL1, SNRPPN, and ZIM2 in bEPSCs. (C) Immunostaining detection of H3K27me3 foci in female bEPSCES. The male bEPSCES were the negative control. (Scale bar, 50 μm.) (D) PCA of global gene expression (RNA-seq) of bEPSCs, bovine-primed ESCs, and BFFs. (E and F) Expression of pluripotency genes, lineage genes, and DNA methylation genes in bEPSCs, pEPSCs, and BFFs. Bovine-primed ESCs, n = 10; BFFs, n = 2; bEPSCsiPS-Q36, n = 2; bEPSCsES-A6, n = 2; n represents the number of biologically independent samples. (G) PCA of global gene expression (RNA-seq) of EPSCs and bovine preimplantation embryos (GSE59186) (60). Two replicates in each sample were used. (H) Expression levels of all annotated bovine histone genes in bovine-primed ESCs, EPSCs, and BFFs.