An active PI3K/AKT pathway is required for oncogenic ETS, but not KRAS, to induce prostate cell migration. (A) An immunoblot shows the levels of pAKT, pMEK (activator of ERK), or tubulin (control) after LY294002 (20 μM; 24 h) or ZSTK474 (2 μM; 24 h) treatment in RWPE-ERG or RWPE-KRAS cells. (B) A transwell assay measured cell migration of RWPE prostate cells with or without ERG and KRAS overexpression and in the presence or absence of the PI3K inhibitors LY294002 (20 μM) or ZSTK474 (2 μM). The number of migrated cells is shown as the mean and SEM of six biological replicates (except for ZSTK474 treated cells which have three replicates) relative to RWPE-empty vector. (C) A transwell assay, as in (A), tested the role of PI3K inhibition on ETV1 and ETV5 expressing RWPE cells and shows the mean and SEM of three biological replicates. (D) Results of the scratch assay performed in the presence or absence of LY294002 (20 μM) and AKT inhibitor VIII (10 μM) in RWPE-ERG (Grey bar) and RWPE-KRAS (white bar) cells. The percentage of scratch filled is shown as the mean and SEM of three biological replicates (each mean of three technical replicates) relative to no treatment. P-values are calculated by t test: * < 0.05, ** < 0.005, *** < 0.0005, unmarked > 0.05.