Fig. 3.

Akt2 accounts for most of the insulin-stimulated Akt protein kinase activity in cultured 3T3-L1 adipocytes. The 3T3-L1 adipocytes were transfected with siRNAs (40 nmol of siRNA duplexes/5 × 10⁶ cells) by electroporation, reseeded for 42 h, and serum-starved for 6 h before the treatment with insulin for 15 min at 37°C. Protein (50 μg) of total cell lysate was used for detection of Akt1 and phospho-Thr-308/309 Akt. (A) Protein (25 μg) was used for detection of Akt2, nonmuscle myosin IIB, and Acrp30. (B) Quantification of Akt1 and Akt2 protein levels, respectively. (C) Quantifications of Akt Thr-308/309 phosphorylation. Data are presented as a mean ± SD of three independent experiments.