Fig. 2: Combination treatment induces apoptosis
a PANC-1 cells were seeded in triplicate in 12-well plates at a cell seeding density of 3×104 cells/cm2 and left to attach overnight. Cells were treated with 100 μM gemcitabine for 24 h and/or 100 ng/ml TRAIL for 6 h. The viability of the cells was assessed by trypan blue exclusion assay. Quadruplicate cell counts were used to calculate each cell density. These were performed for three independently seeded wells and percentage viability was determined. b A total of 1×106 PANC-1 cells were treated with 100 μM gemcitabine for 24 h and/or 100 ng/ml TRAIL for 6 h. Induction of early apoptosis in PANC-1 cells was assessed using flow cytometry following staining with FITC Annexin V. The data represent means ± SEM of three experiments performed in triplicate. c, d PANC-1 cells were seeded in triplicate in 12-well plates at a cell seeding density of 3×104 cells/cm2 and left to attach overnight. Cells were treated with 100 μM gemcitabine for 24 h and/or 100 ng/ml TRAIL for 6 h and monitored by time-lapse microscopy. c The appearance of a pre-apoptotic morphology was scored and the % apoptotic cells after 24 h determined. The data are the means ± SEM from three independent experiments. d Phase contrast microscopy images of cells treated as indicated. a–c All experiments were repeated three times and data are provided as means ± SEM (one representative experiment is shown). P-values were calculated using Student’s t-test to determine the statistical significance of the difference between cells treated with 100 μM gemcitabine and cells treated with 100 μM gemcitabine plus 100 ng/ml TRAIL (*P < 0.05, **P < 0.01, ***P < 0.001)
