Figure 3
From: Inhibition of AKT survival pathway by a small molecule inhibitor in human endometrial cancer cells
Effect of API-59CJ-OMe on AKT kinase activity and phosphorylation of other kinases. (A) After treatment with API-59CJ-OMe, cells were harvested and cell lysates separated by 10% SDS–PAGE. Gels were analysed with phospho-specific antibodies to PDK1 (Ser 241), AKT (Ser 473 or Thr 308), ERK1/2 (Thr 202/Tyr 204), JNK (Thr 183/ Tyr 185), SGK (Ser 78), PKCζ/λ (Thr 410/403), and PKCα/β (Thr 638/641). The same lysates were immunoprecipitated with anti-AKT or anti-ERK antibody, and kinase assays performed with GSK-3 as a substrate for AKT and Elk-1 as a substrate for ERK. AKT (IP) represents the immunoprecipitation control: lysates were immunoprecipitated with mouse monoclonal anti-AKT antibody, separated by SDS–PAGE, and stained with rabbit polyclonal anti-AKT antibody. Phospho-specific antibodies to GSK-3 α/β (Ser 21/9) and Elk-1 (Ser 383) were used for phosphorylated protein detection.
