Effects of Cav-1 levels on the phosphorylation state of ERK and AKT.

<p>Control, over-Cav-1 cells and shCav-1 cells were seeded in 60-mm dishes (at a density of 12×10⁴). A) At the indicated time points, cells were harvested and protein homogenates western blotted for pAKT, AKT, pERK, ERK, pCav-1, Cav-1 and tubulin at the indicated time points. Protein bands corresponding to pAKT and pERK levels (as detected at 48 hours) were quantified by densitometry after normalization with respect to tubulin (n = 3). **, P<0.001; ***, P<0.0001. B) 24 hours after seeding, mock and over-Cav-1 cells were treated with 15 µM PD09859 (in the case of RD cell line) or 10 µM LY294002 (in the case of RH30 cell line) for up to 24 hours. Cell proliferation was then evaluated by Crystal violet assay. Histograms represent means ± SD of absorbance (n = 4). *, P<0.05; **, P<0.001; ***, P<0.0001.