Figure 11. Effect of A66 + akti1/2 and wortmannin + akti1/2 on lipolysis.

Adipocytes were pre-incubated without (blue) (a–f), with 10 µM A66 + 20 µM akti1/2 (a–c) (green and brown), or with 0.5 µM wortmannin + 20 µM akti1/2 (d–f) (green and brown) for 30 min, when the indicated concentration of insulin was added and cells incubated for 15 min before incubation with 10 nM isoproterenol (iso) for another 10 min. Cells were then separated from medium and the phosphorylation of PKB at Ser⁴⁷³ (a and d) or Thr³⁰⁸ (b and e) was determined in the cells, and glycerol released to the medium was determined (c and f) (n = 5). Indicated are P-values for the effect of indicated inhibitors in isoproterenol-stimulated cells (c and f). Representative immunoblots of one experiment are shown [blots in (a) and (b), and in (d) and (e), respectively, are from the same experiment, hence identical β-tubulin blots].