Fig. 7

ETV4 downregulates the CDKN1A promoter also through the modulation of p53 levels. a Dual luciferase reporter assay in PNT1A and RWPE cells transiently transfected with a vector containing firefly luciferase downstream the 20 bp human CDKN1A p53-binding site (p53 BS) (top panel). The bar diagram shows the relative luciferase activity (Firefly/Renilla ratio) from cells transfected with either an ETV4-expressing vector (+ETV4, grey bars) or a p53-expressing vector (+p53, striped bars), and normalized to those transfected with an empty vector (CTL, black bar). Data represent average and standard error of triplicate measurements of 3 independent experiments. b Representative western blots of p53 in human RWPE cells transfected with either an empty vector (CTL) or an ETV4-expressing vector (+ETV4). Beta actin is used as loading control. c Bar diagram of p53 protein levels measured by western blot analysis in human RWPE cells transfected with an ETV4-expressing vector (+ETV4) normalized to those transfected with an empty vector (CTL). Average and standard error values of the relative quantification are shown. Seven independent experiments have been performed. d Representative western blots of p53 protein in the 3 prostate lobes of wild-type (WT) and of ETV4 transgenic mice (lines ETV4 A and ETV4 B); beta-actin was used as loading control. The relative densitometric quantifications are shown for each lane. e Bar diagram of p53 protein levels measured by western blot analysis in the 3 prostate lobes of ETV4 transgenic mice (lines ETV4 A and ETV4 B) normalized to wild-type mice (WT). Average and standard error values of the relative quantification are shown; n: number of mice. *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001