Fig. 2.

Serum starvation does not affect insulin-stimulated Akt or AS160 phosphorylation. Differentiated L6 myotubes were incubated 3 h in serum-replete (control) or serum-free medium (serum-starved) and further incubated at 37°C for 20 min in the presence or absence of 100 nM insulin before extraction with ice-cold lysis buffer. Cell lysates were subjected to SDS/PAGE and Western blot analysis. A: P-Akt S473 and Akt (n = 3/group). B: P-Akt T308 and Akt (n = 3/group). C: phosphorylated Akt substrate and AS160 (n = 3/group). Data represent means ± SE. *Significant difference due to serum starvation (P < 0.05). For A–C, there is a main effect for insulin (P < 0.05).