Fig. 2. IGF-I induces phosphorylation of Akt on S473. Serum-deprived SHEP cells are exposed to DMEM (B and D) or DMEM + 10 nM IGF-I (A and C). After 0, 5, 15, 30, 45, and 60 min, cell lysates are collected and run on a 12.5% SDS-PAGE gel, transferred to nitrocellulose, and Western immunoblotted for either P-S473-Akt (A and B) or Akt (C and D).