Fig. 3. Total Akt and phosphorylated Akt in whole cell lysates determined by SDS PAGE on 10% gels and immunoblotting. Cells were stimulated with 10 ng/ml EGF for 10 minutes or 10 nM mut C uPA. (A) uPA activated Akt phosphorylation. (B) Inhibition of EGF-stimulated Akt phosphorylation by 1-10 µM LY 294002 (pre-treatment 1 hour). Relative band intensities represent the ratio of phospho-Akt to total Akt (band intensity in unstimulated cells, 1.0). The results are representative of three experiments.