Figure 2

Differential and isoform specific activation of AKT1, AKT2 and AKT3. (A) Comparison of AKT isoform expression in HCC cell lines. Exponentially growing cells were seeded into 10 cm dishes and allowed to attach for 24 h. Cells were then lysed and analyzed by Western blot. One representative experiment out of three is shown. (B) HCC cells were treated over 24 h with the indicated concentration of RAD001. AKT isoform specific in vitro kinase assays were performed following quantitative AKT isoform immunoprecipitation, based on the same cell lysates as shown in Figure 1A and B. GSK3αbgr; fusion protein was used as an AKT substrate and phosphorylation at (S9/21) detected by Western blot. Columns, mean of three independent experiments; bars, SD. * p < 0,05; # p < 0,01.