Figure 3.

TET3-dependent epigenetic regulation of the DUSP family

(A and B) mRNA and protein expression of genes of interest in HTR-8/SVneo cells transfected siTET3 or siCon was measured using quantitative real-time PCR and western blot, respectively. Total RNA and protein were isolated at 48 h after transfection (n = 3). (C–E) Sequences of critical transcription regulatory regions of DUSP2, DUSP4, and DUSP5. The number indicates the position of the nucleotide relative to the transcription initiation site. Differentially methylated cytosine residues are highlighted in red. HTR-8/SVneo cells were transfected with siTET3 or siCon. MethylTarget sequencing analysis was performed at 48 h post-transfection. The methylation levels of the CpG sites in the DUSP2, DUSP4, and DUSP5 genes are shown (F–H) (n = 3). Genomic DNA was extracted from human placental tissues (n = 4 per group) and analyzed using MethylTarget sequencing. The promoter methylation levels of the CpG sites in the DUSP2, DUSP4, and DUSP5 genes are shown (I–K). The total promoter methylation levels of the CpG sites in the DUSP2, DUSP4, and DUSP5 genes are presented (L–N). Error bars indicate mean ± SEM. ∗p < 0.05 and ∗∗p < 0.01.