Figure 5.

Effect of LY294002 on Akt, p27^KIP1, and cell cycle profile. (A) Western blot analysis of drug-treated cells. Actively growing U87 cells (Left) or DLD1 cells (Right) were treated with either dimethyl sulfoxide (control) or LY294002 (20 μM) for 24 hr. Cell lysates (50 μg each) were examined by Western blot analysis with phosphorylation status specific or regular antibodies against Akt, respectively. A duplicated blot was also examined by Western blot analysis with anti-p27 antibody. (B) Fluorescence-activated cell sorting analysis of cell cycle profile of drug-treated cells. Actively growing U87 cells (Upper) or DLD1 cells (Lower) were treated with either dimethyl sulfoxide (control) or LY294002 (20 μM) for 24 hr. Cells were harvested and examined by fluorescence-activated cell sorting analysis for cell cycle distribution as described in the legend of Fig. 3A.