Fig. 4

Sin3a recruits Tet1 and facilitates the hydroxymethylation of epithelial gene promoters. A Co-IP analysis for the interaction of endogenous SIN3A and TET1 at Day 5 of reprogramming. 10% of total lysates were used for input. Rabbit IgG antibody as the negative control. B–E ChIP-qPCR analysis (n = 4) of Sin3a enrichment at the promoter of epithelial markers (Cldn3 (B), Pkp1 (C), Krt8 (D), and E-cadherin (E)) at Day 5. The samples were normalized to input DNA. F–I ChIP-qPCR analysis of Tet1 occupation at the promoter of epithelial genes at Day 5. J hMeDIP-qPCR was employed to detect the 5hmC level at the promoter of epithelial markers after Sin3a knockdown. *P < 0.05, **P < 0.01, and ***P < 0.001 versus the control group. K qRT-PCR analysis of Tet1 expression at Day 5 and Day 8 of reprogramming. MEFs were infected with scramble (shCtrl) or shSin3a (shTet1-1 or shTet1-2) lentivirus. L Representative images of cell morphology at Day 5 of reprogramming. The black arrow indicates the representative epithelial morphogenesis. M, N qRT-PCR analysis of the expression changes of epithelial genes (M) and mesenchymal genes (N) at Day 5 and Day 8 of reprogramming after Tet1 knockdown. Significance was estimated by student’s unpaired t test (n = 3 independent experiments). *P < 0.05, **P < 0.01, and ***P < 0.001 versus the control group at Day 5. ^#P < 0.05, ^##P < 0.01, and ^###P < 0.001 versus the control group at Day 8. Error bars represent SD