Fig. 3.

Effect of Ser129Ala mutation on the Thr308 phosphorylation in response to different cellular conditions. HEK 293T cells were transfected with w.t. Akt or Ser129Ala Akt mutant for 24 h and treated as indicated; 10 μg of total proteins from cell lysates were analyzed by Western blot. a After transfection, cells were serum-deprived for 12 h, then treated with 100 ng/ml IGF-1 for the indicated times. b Cells were treated with 0.5 μM okadaic acid (OA) where indicated for the last 1 h of transfection. c Cells were treated with 25 μM LY294002 (LY) where indicated for the last 2 h of transfection