Figure 3. Analysis of transcriptional control in MCM7 gene in MCF7 cells in response to CDK4/6 inhibitor. (A) Semi-quantitative RT-PCR analysis of MCM7 and GAPDH mRNA expression. (B) Western blotting analysis of pRB, phospho RB (Ser 780), MCM7 and loading control lamin b. (C) Analysis of BrdU incorporation by flow cytometric analysis. (D) Schematic representation of RB/E2F-biding nucleotide sequences on MCM7 promoter. (E) Identification pRB association on RB/E2F consensus binding site I on MCM7 promoter (ChIP assay). (F) RB ChIP on RB/E2F consensus binding site II of MCM7 promoter in response to DMSO or PD 0332991 in MCF7 cells. (G) RB ChIP on RB/E2F consensus binding site III of MCM7 promoter in response to DMSO or PD 0332991 in MCF7 cells. (H) Identification acetylated histone 4 association on RB/E2F binding site I on MCM7 promoter (ChIP assay). (I) MCM7 promoter analysis (luciferase assay). Each data point is a mean ± SD from three or more independent experiments. p < 0.05 were considered as significant.