Fig. 6

ETV4 downregulates CDKN1A promoter regardless of ETV4-binding sites. a Diagram of the human CDKN1A promoter (top) and of the vectors (BS-AB) in which firefly luciferase expression is driven by the 1656 bp region upstream the CDKN1A TSS (bottom). ETV4 BSs are indicated by a diamond and p53 BSs are indicated by a circle. Four different variants of the BS-AB vector have been used. The BS-AB WT with both wild-type ETV4 BS-A and BS-B. In the other 3 vectors, one or both of this ETV4 BS have been mutated; the mutated sites are in black the normal sites are in white. b Quantification of dual luciferase reporter assay in PC3 cells transiently transfected with the indicated four BS-AB vectors (see above). The bar diagram shows the relative luciferase activity (Firefly/Renilla ratio) from cells transduced with one of 2 anti-ETV4 shRNA (shETV4a and shETV4b) normalized to those transduced with an irrelevant shRNA (CTL). Data represent average and standard error of triplicate measurement of 3 independent experiments. c Quantification of dual luciferase reporter assay in RWPE cells transiently transfected with the indicated four BS-AB vectors (see above). The bar diagram shows the relative luciferase activity (Firefly/Renilla ratio) from cells transfected with an ETV4-expressing vector (+ETV4) normalized to those transfected with an empty vector (CTL). Data represent average and standard error of triplicate measurement of 3 independent experiments. *P ≤ 0.05, **P ≤ 0.01