Effect of HCV infection on insulin-induced phosphorylation of Akt and GSK3β. Huh7.5.1 cells were seeded in a 24-well plate and subjected to serum-free incubation for 20 h, followed by insulin stimulation for the indicated time (0, 15, 30, and 60 min). Western blotting with indicated antibodies revealed a time-dependent change in the phosphorylation of Akt (a) and GSK3β (b). Furthermore, Huh7.5.1 cells and HCV-infected cells were stimulated with 100 nM insulin for 30 min or left untreated. Extracted proteins were analyzed with antibodies against pAkt, Akt, pGSK3β, and GSK3β (c).