Fig. 5
Supraphysiological β-estradiol (E2) induces the expression of ten-eleven translocation 2 (TET2) and alters the levels of global DNA 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) in mouse parthenogenetic embryonic stem cells (pESCs). A The derivation process and AP staining of pESCs; P2: passage 2; P9: passage 9; P16: passage 16. Representative image of alkaline phosphatase (ALP) staining in pESCs, Octamer-Binding Transcription Factor 4 (OCT4) and sex-determining region Y-box 2 (SOX2) expression in pESCs-blue fluorescence with 4,6-diamino-2-phenyl indole (DAPI) nuclear staining and green fluorescence with Dylight 488 staining. B Representative immunofluorescence images of pESCs as shown with 5mC and 5hmC (red fluorescent) and nuclear DAPI staining (blue fluorescence) within each group. C Quantification of average fluorescent intensities for 5mC and 5hmC staining within each group (n = 6). D, E The percentage of methylated DNA, hydroxymethylated DNA and 5-hmC/5mC ratio were detected by Enzyme-Linked Immunosorbent Assay (ELISA) kit (n = 6). F Analysis of methylation levels at cytosine preceding a guanine base (CpG) sites within the differentially methylated region (DMR) in the mesoderm-specific transcript homologue (Mest) promoter, evaluated using bisulfite sequencing PCR (BSP, n = 9). Data are expressed as the means ± standard deviation (SD), **P < 0.01, ***P < 0.001. Scale bar = 100 μm (unpaired two-tailed t test)