DNA Methyltransferase Controls Stem Cell Aging by Regulating BMI1 and EZH2 through MicroRNAs (original) (raw)
Figure 2
DNMT inhibition induced cellular senescence.
(a) hUCB-MSCs were treated with the DNMT inhibitor 5-AzaC for 7 days. DNMT inhibition by 5-AzaC induced cellular senescence, as shown by SA β-gal staining. (b) After 5-AzaC treatment for 1, 2 and 3 days, an MTT assay was performed. (c–d) 5-AzaC increased p16INK4A and p21WAF1/Cip1 and decreased DNMT1, DNMT3A and DNMT3B, as shown by real-time qPCR analysis (c) and western blot analysis (d). 5-AzaC treatment for 1, 3, 5 and 7 days induced cellular senescence of hUCB-MSCs, as shown by SA β-gal staining (d). (e) After a 2 day treatment with 5-AzaC, FACS analysis was performed, as described in the Materials and Methods section. 5-AzaC treatment induced G1 phase cell cycle arrest in a dose-dependent manner. CDK2 and CDK4 expression levels were confirmed by western blot analysis.