Figure 1

Hes1 inhibits δ-catenin transcription in prostate cancer cells. (A) A schematic graph showing predicted binding regions of Hes1, E2F1, and p53 on δ-catenin promoter. Numbers on top indicate the sequence location in base pairs from the transcription start site. (B) Hes1 is a negative regulator of δ-catenin transcription. E2F1, WT-Hes1 and DN-Hes1 expression vectors together with two human δ-catenin-luciferase reporter vectors (BK1 and BK5) and a control vector (pGL3-Basic), were co-transfected into PC3 and CWR22-Rv1 cells (0.2 μg/well for each vector in 12-well plates) as indicated. Hes1 blocked δ-catenin-luciferase activity completely, which was induced by E2F1. DN-Hes1 did not block E2F1 induced δ-catenin-luciferase activity, regardless of cell types and reporter vectors used. (C and D) Hes1 inhibited δ-catenin-luciferase activity (BK5 reporter vector) in a dose-dependent manner. PC3 cells were co-transfected with pcDNA-flag-WT-Hes1 and E2F1 vectors. (C) E2F1 plasmid was used at 0.2 μg/well and pcDNA-flag-WT-Hes1 plasmid was used for co-transfection at 0.02 μg, 0.04 μg, 0.1 μg and 0.2 μg per well separately in 12-well plates. (D) pcDNA-flag-WT-Hes1 plasmid was used at 0.05 μg/well and E2F1 plasmid was used for co-transfection at 0.3 μg, 0.2 μg, 0.1 μg and 0.05 μg per well separately in 12-well plates. The data was representative of three independent experiments.