Figure 1.

MPL inhibits apoptosis through down regulating caspases. A. Western blot of lysates prepared from OVCAR-3 and A2780 cells treated with MPL (0, 5, 10, 25 μM for 48 and 72 h) were analysed for the expression of caspase-3 and caspase-8 proteins. The house-keeping gene (GAPDH) was used to confirm similar protein loading and blot transfer. A representative from three independent experiments has been shown. B. The impact of MPL (0, 10 and 25 μM) on caspase-3 and -8 activities was assessed in MPL-treated OVCAR-3 cells for the indicated time points (48-72 h). Each concentration was tested in replications of 4 and each experiment was repeated twice. For statistical comparisons, each drug treated group was compared with the control group using Student’s t-test. C. Cell viability estimated by SRB assay in OVCAR-3 and A2780 cell lines after treatment with MPL (0, 10, 25 μM) 30 min. after pan-caspase inhibitor z-VAD-fmk (5 μM) up to 72 h. Each concentration was tested in replications of 8 and each experiment was repeated twice. Data represent mean ± SEM from two independent experiments combined. D. Cells treated with MPL (0, 10 and 25 μM) for the indicated time points (24, 48 and 72 h), stained with Annexin-V/7-AAD and cell death rates analysed by FACS. Experiment repeated twice for each concentration and time points. Data represent mean ± SEM from two independent experiments combined. E. DNA ladders were detected on 1.5% of agarose gel after OVCAR-3 and A2780 cells were treated with 25 μM MPL for 48 h. A representative from three independent experiments is shown.