Figure 2.

Insulin-stimulated phosphorylation of PKB/Akt and its downstream substrates is impaired by nelfinavir (Nelf). 3T3-L1 adipocytes were treated without or with 30 μm nelfinavir for 18 h and then washed and stimulated for 10 min with insulin. Cell lysates and Western blot analysis were performed as described in Materials and Methods, and membranes were blotted with antibody against PAS (A); anti-pSer473-Akt/PKB antibody and anti-pSer9-GSK3 antibody or antibodies against total Akt/PKB and GSK3 (B); or anti-PAS and anti-AS160 antibodies (C). Shown are blots representative of three to five independent experiments. Anti-PAS blots in A are shown after both long and short exposure to depict various phosphorylated PAS below signal saturation conditions. Cont, Control; pS, pSer 9.