Figure 5.

Statin-induced Akt phosphorylation is abolished by treatment with LY294002 but not with 25HC. (A) 13A/PS cells were incubated for varying times (0–4 h) in medium B in the absence or presence of compactin (5 μM) and were coincubated in the absence or presence of LY294002 (20 μM) or hydrogen peroxide (500 μM). (B and C) 13A/PS cells were incubated for 30 min in medium B in the presence of compactin (5 μM) and were coincubated in the absence or presence of LY294002 (20 μM) or 25HC (1 μg/ml). Panel C shows the relative intensity of the P-Akt band relative to the total Akt band (means + SEM) from three separate experiments. There is no error bar in the compactin only control in C, because this condition was set at 1 for all three separate experiments. Whole cell lysates (containing 80 μg protein) were subjected to 10% SDS-PAGE and immunoblot analysis with anti-Phospho-Akt (Ser473; P-Akt) or anti-Total Akt as described in Materials and Methods. Relative intensity of the P-Akt band relative to the total Akt band was quantified by densitometry and presented relative to control in each panel.