Endocrine-Related Cancer

E2F1 binding to the hPOMC promoter. (A) Results of EMSA performed with indicated probes and consensus competitors. No competitor is indicated by -. (B) EMSA and supershift-EMSA were performed with nuclear extracts prepared from DMS79 cells using probe #5, E2F consensus competitor, and indicated antibodies to E2F molecules. (C) Results of luciferase assays performed using the indicated reporter plasmids constructed with hPOMC promoter fragments (#5, #6, and #7) and the minimal promoter shown in Fig. 2B. Reporter plasmids were co-transfected with E2F1 or E2F3 expression plasmids or negative control empty vector (Vector). (D, E, F) ChIP assays were performed with anti-E2F1 (E2F1), anti-acetyl histone H4 (AcH4), or negative control (IgG) using primer sets shown in D (a, b, c). The analyzed ChIP regions are from DMS79 (E) and COLO320 cells (F). EMSA and ChIP results are each representative of triplicate independent experiments (A, B, E, F) and luciferase results are representative of four independent experiments, each depicted as mean ± s.e. of triplicate samples (C).