VEGF triggers time- and dose-dependent protein expression in MM1s cells, which is specifically inhibited by GW654652. (A) VEGF triggers dose-dependent up-regulation of Mcl-1 expression in MM1s cells. MM1s cells were starved overnight in RPMI 1640 supplemented with 0.5% FBS, followed by culture in the presence or absence of the indicated doses of VEGF for 6 hours. Cell lysates (30 μg) were investigated by Western blot analysis with indicated antisera. Actin was used as a loading control. (B) VEGF triggers time-dependent modulation of protein expression in MM1s cells. MM1s cells were cultured overnight in RPMI 1640 supplemented with 0.5% FBS, followed by culture with VEGF (50 ng/mL) for 6 hours. Cell lysates (30 μg) were studied by Western blot analysis with indicated antisera. Actin served as a loading control. (C) VEGF-triggered Mcl-1 up-regulation is inhibited by GW654652. After overnight starvation followed by addition of the indicated doses of GW654652 (1 hour), cells were cultured in presence or absence of 50 ng/mL VEGF (6 hours). Cells lysates (30 μg) were analyzed by Western blot analysis. Shown is 1 representative experiment of 3.