DNA Methyltransferase Controls Stem Cell Aging by Regulating BMI1 and EZH2 through MicroRNAs (original) (raw)
Figure 4
DNMT inhibition modified histone marks, transcriptional enzymes and the CpG island methylation status in the CDKi promoter regions.
(a–b) After treatment with 5-AzaC for 5 days, methyl-specific PCR was performed. (a) Schematic diagrams indicate locations of each primer on CDKi promoter regions. (b) Methyl-specific PCR was performed as described in the Materials and Methods section. M: methyl primer, U: unmethyl primer. (c–f) After treatment with 5-AzaC for 3 days, ChIP analysis was performed using antibodies targeting the indicated protein (AcetylH3, AcetylH4, H3K4Me3, H3K9Me3, H3K27Me3, PolII and EZH2). (c) Schematic diagrams indicate the locations of each primer on genomic DNA. (d–f) Fold enrichment of indicated proteins on the promoters of p16INK4A and p21WAF1/Cip1 were investigated by real-time PCR.