TRAIL-induced apoptosis of hepatocellular carcinoma cells is augmented by targeted therapies

Figure 5 TRAIL-induced apoptosis in Huh7 cells after targeted therapy approaches and knock-down of BCL-x_L and MCL-1. A: Huh7 cells were transfected with siRNAs (40 nmol/L) specific for MCL-1 and BCL-x_L either alone or in combination. SiGFP was used as control. Whole cell lysates were prepared 24 h after transfection. MCL-1 and BCL-x_L expression was analyzed by Western blotting. α-Tubulin expression was used to control equal loading; B: 24 h after siRNA transfection, cells were treated for 48 h with 100 ng/mL TRAIL (+ 1 μg/mL Enhancer); C: 20 μmol/L SP600125 (left panel) or 10 μmol/L LY294002 (+ 0.2% DMSO as vehicle, right panel), either alone or in combination with 100 ng/mL TRAIL (+ 1 μg/mL Enhancer). Cells were harvested on day two after treatment and analyzed for apoptosis induction by flow cytometry. Assays were performed in triplicate and are representative of at least two independent experiments. Values are expressed as mean ± SD. NS: Not significant. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001.