Fig. 4

DU145 cells adhesion, invasion and intracellular activated pathways upon exposure to LO145 or LOR80. a DU145 adhesion, expressed as % of adherent cells, upon exposure to LO145 or LOR80 vs vehicle (considered as 100% control) and evaluated both in absence or presence of vitronectin (VN) for either 30 or 60 min. Stat. significances are expressed on vehicle. b DU145 adhesion measured as above, on VN after 60′ of exposure to LO145 or LOR80, untreated or pre-treated for 45′ RT with anti-αV-integrin monoclonal blocking antibody (anti-αV Ab) and indicated as LO145^αv and LOR80^αv, respectively; as negative control cells were treated by the anti-αV Ab alone. (LO145^αv/ LO145)**; (LOR80^αv/ LOR80)***; (LOR80/ vehicle) ***. c Western blotting analysis of pFAK, FAK, pAKT, AKT and γ-tubulin on cell lysates from DU145 untreated (CTR), or treated with LO145 or LOR80, alone or pre-exposed to anti-αV Ab (see above). pFAK and pAKT quantification is also reported. d Adhesion of DU145 measured as above, untreated (considered as 100%) or pretreated with the AKT inhibitor GDC-0068 for 16 h, were exposed to vehicle, LO145 or LOR80. (GDC-0068/ vehicle)***; (LOR80/ vehicle) **; (GDC-0068+ LOR80/ LOR80)* (e) DU145 invasion, expressed as % of invading cells, upon 16 h exposure to LOR80 treated with anti-αV Ab (LOR80^av) or control Ab (LOR80^{CTR Ab}) vs vehicle (considered as 100% control); as negative control cells were treated by CTR Ab alone (anti-pAKT antibody ineffective on non-permeabilized intact cells); (LOR80/ vehicle)***; (LOR80^{CTR Ab}/vehicle)***; (LOR80^av/LOR80)** (f) Invasion of DU145 measured as above, untreated (considered as 100%), or pretreated with the AKT inhibitor GDC-0068 for 6 h and exposed to vehicle, LO145 or LOR80;(GDC-0068/ vehicle)***; (LOR80/ vehicle)***; (GDC-0068 + LOR80/ LOR80)***. Results are representative of a single experiment and several experiments yielded similar results (b), or are the average of at least two experiments (a; d; e; f)