Figure 4

Effect of methylation of site CGCG (−695~-692) on MYCT1 promoter banding to c-Myc in Hep-2 cells. A. Binding ability of site CGCG (−695~-692) to c-Myc. End-labeled M.SssI-treated methylated and unmethylated MYCT1 promoter region probes containing CGCG (−695~-692) were mixed with nuclear proteins from Hep2 cells and separated on a 6% polyacrylamide gel. Lane 1, end-labeled M.SssI-treated methylated free probe; Lane 2, end-labeled M.SssI-treated methylated probe mixed with nuclear proteins; Lane 3,4, the unlabeled M.SssI-treated methylated wild-type competitor DNA was mixed with the nuclear proteins before adding end-labeled probe; Lane 5, c-Myc antibody was mixed with the nuclear proteins before adding end-labeled probe; B. Binding of E-box B to c-Myc in vivo detected by ChIP. The percentage of the MYCT1 promoter occupied by c-Myc was significantly different in Hep-2 cells of present or absent 5-zaz..** p < 0.01.