Figure 4

The PI3K pathway can alter the expression of cell migration genes via ETS/AP-1 sites in oncogenic ETS overexpressing cells. mRNA expression of (A) ARHGAP29 or (B) SMAD3, in the presence and absence of PI3K inhibitor (LY294002, 20 μM), in RWPE-ERG and RWPE-KRAS cells was measured by qRT-PCR and compared to RWPE cells. Mean and SEM of seven biological replicates are shown. (C) Firefly luciferase activity from a vector with the indicated sequences (3 copies of neighboring ETS and AP-1 binding sequences or versions of the same with point mutations) is shown relative to Renilla luciferase from a control vector transfected in RWPE cells. The ERK pathway is inhibited by UO126 where indicated. Mean and SEM of six biological replicates (each mean of two technical replicates) are shown. Luciferase reporter activity measured as in (C) is shown in (D) RWPE-ERG, or (E) RWPE-KRAS cells as activity in LY294002 treated cells (20 μM) relative to untreated. P-values are calculated by t test: n.s > 0.10, * < 0.05.