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. 2016 Aug 4;6:30922. doi: 10.1038/srep30922

Figure 7. DNMT1 represses p53 by binding to its promoter locus.

Figure 7

ChIP assay was used to further confirm DNMT1 binding activity in all seven groups. Chromatin was prepared from treated CPCs and immunoprecipitated using DNMT1 antibody. DNA sequence fragments from the p53 promoter onto which DNMT1 was recruited in all groups were amplified by PCR using specific primers. Data were obtained from three independent experiments under the same experimental conditions. Input, input chromatin prior to immunoprecipitation. α-IgG, immunoprecipitation with normal rabbit IgG. α-DNMT1, immunoprecipitation with DNMT1 antibody. gDNA, amplification of genomic DNA containing the p53 promoter sequence. No Ab, immunoprecipitation without antibody. H0, normoxia. H0+24 h, normoxia + oxygen–serum deprivation for 24 hours. H6, HP for 6 hours. H6+24 h, HP for 6 hours + oxygen–serum deprivation for 24 hours. H6+AZA+24 h, HP for 6 hours with 5-azadc (a DNA methyltransferases inhibitor) + oxygen–serum deprivation for 24 hours. H6+siD1+24 h, HP for 6 hours with siRNA-DNMT1 + oxygen–serum deprivation for 24 hours. H6+AZA+siD1+24 h, HP for 6 hours with 5-azadc and siRNA-DNMT1 + oxygen–serum deprivation for 24 hours.