Fig. 5

c-Src-mediated DMAP1 phosphorylation blocks DMAP1-mediated DNA methylation. a, SW1990 cells expressed with the indicated plasmids were synchronized in mitosis (M) by nocodazole (200 nM) treatment for 16 h after releasing thymidine double block for 8 h. DNA methylation levels of promoters and CpG islands or CpG islands shores were presented as ratio of methylated reads to unmethylated reads. The values represent from 2 repeated samples. b, SW1990 cells expressed with the indicated plasmids were synchronized in mitosis (M) by nocodazole (200 nM) treatment. DNA methylation profile of the promoter region (TSS ±1 kb) of BCL2L1. The values represent from 2 repeated samples. c, SW1990 and PANC-1 cells expressed with the indicated plasmids were synchronized in mitosis (M) by nocodazole (200 nM) treatment. 5-mc levels were examined by bisulfite sequencing using primers covering TAp73 binding site of BCL2L1 gene promoter region. d-g, Cells were synchronized in interphase (I) by thymidine (2 mM) double block or were synchronized in mitosis (M) by nocodazole (200 nM) treatment. d, SW1990 cells with depleted DMAP1, and reconstituted expression of WT rDMAP1 or rDMAP1 Y246F were transfected with plasmid for expression of TAp73 shRNA. Relative mRNA level was analyzed by real-time PCR. e, ChIP analyses were performed in SW1990 cells. The primers covering TAp73 binding site of BCL2L1 gene promoter region were used for the real-time PCR. f, SW1990 cells were transfected with plasmid for expression of TAp73 shRNA. ChIP analyses were performed. The primers covering TAp73 binding site of BCL2L1 gene promoter region were used for the real-time PCR. g, SW1990 cells were expressed with the indicated plasmids. ChIP analyses were performed. The primers covering TAp73 binding site of BCL2L1 gene promoter region were used for the real-time PCR. The y axis shows the value normalized to the input. The values represent mean ± s.e.m. of three independent experiments;*represents p < 0.05 and **represents p < 0.01 between indicated groups