The phosphor-AKT and phosphor-S6 of the SMM103 melanoma cells treated with sirolimus were detected by western blot, SMM103 cells were exposed to various concentrations of sirolimus for 72 hours, sirolimus inhibited the phosphor-S6 at Ser240/244 and activated the phosphor-AKT at Ser473, starting at doses as low as 0.1nM (Figure 4B, left). The gene discussed is AKT1; the disease is melanoma.