Fig. 3. Kaiso binds to the GR promoter sequence via a methyl-CpG-dependent mechanism. (A) Schematic diagram of the GR proximal promoter. The two main alternative exons are labeled 1B and 1C. Relative to the translation start site, the P1 primer set amplifies the 283 bp region from −4046 to 3764, the P2 primer set amplifies the 209 bp region from −3549 to −3339, the P3 primer set amplifies the 204 bp region from −2868 to −2665, and the P4 primer set amplifies the 298 bp region from −2294 to −1997. (B) MCF-7 cells were induced to overexpress FLAG-fused Kaiso. Chromatin was immunoprecipitated using an antibody against FLAG or (C) Kaiso in T47D cells. Anti-IgG antibody was used as a negative control. Sets of primers were used to amplify the various GR promoter regions from immunoprecipitated chromatin. The GR promoter plasmid was used as a positive control for PCR. (D) T47D cells were treated with 5-AZ, and Kaiso expression was determined by western blotting. (E) ChIP analysis of the various GR promoter regions in T47D cells. Data shown represent results of two independent experiments.