Effects of combined treatment on cell cycle distribution and apoptosis in three HCC cell lines. (A) Cell cycle analysis of HCC cell lines after 24h treatment with 2 µM MK-2206, 1 µM AZD6244, 100 nM AZD8055, or the combination of two inhibitors, as indicated. Control cells were treated with DMSO only. Columns: mean of one representative experiment performed in triplicates; bars, SD. (B) HCC cells were seeded into 96-well plates and treated with afore mentioned compounds and concentrations for 24 hours, before apoptosis was measured by Cell Death Detection ELISA. Controls were treated with DMSO. Each data point represents three independent experiments, performed in triplicates; bars: SD. *, p <0.05.