Figure 5.

Role of miR-377 in modulating promoter methylation levels of senescent-associated genes in HSFs. (a) The promoter methylation levels of FoxD3, p53 and UTF1 were analyzed in young HSFs (PD<10) transfected with control mimics or miR-377 mimics through microfluidic PCR and next-generation bisulfite sequencing (data represented as the mean±S.E.M. *P<0.05). (b) The promoter methylation levels of FoxD3, p53 and UTF1 were analyzed in passage-aged HSFs (PD>50) transfected with control inhibitors or miR-377 inhibitors through microfluidic PCR and next-generation bisulfite sequencing (data represented as the mean±S.E.M. *P<0.05). (c) p53 mRNA was detected by RT-qPCR in young HSFs (PD<10) treated with miR-377 mimics together with control cDNA or DNMT1 cDNA as indicated (data represented as the mean±S.E.M. n=3, *P<0.05). (d) DNMT1, p53, and Rb expressions and phosphorylation of Rb were detected by western blot in young HSFs (PD<10) treated with miR-377 mimics together with control cDNA or DNMT1 cDNA as indicated (*P<0.05). Representative data was shown. (e) p53 mRNA was detected by RT-qPCR in passage-aged HSFs (PD>50) treated with miR-377 inhibitors together with control cDNA or DNMT1 cDNA as indicated (data represented as the mean±S.E.M. n=3, *P<0.05). (f) DNMT1, p53, and Rb expressions and phosphorylation of Rb were detected by western blot in passage-aged HSFs (PD>50) treated with miR-377 inhibitors together with control shRNA or DNMT1 shRNA as indicated (*P<0.05). Representative data was shown