Fig. 4

M2I-1 induced caspase-3 activity in the presence of nocodazole. a Cell extracts were prepared from HeLa cells under different drug treatments as indicated. 60 µg/lane of total protein from each sample was loaded for protein separation using a 10% precast SDS-PAGE tris-bis protein gel, and after western blotting the membranes were probed with an anti-caspase-3 antibody (Abcam, ab32351, 1:1000 dilutions), an anti-GAPDH antibody (Thermo Fisher Scientific, MA5-15738, 1:1000 dilutions) which acted as the loading control. Lane 1: HeLa cells treated with 0.5%DMSO (v/v) as control; lane 2: Cells treated with 50 µM M2I-1; lane 3: Cells treated with 60 ng/ml nocodazole; and lane 4: Cells treated with 60 ng/ml nocodazole + 50 µM M2I-1 for 24 h. b Quantitative results comparing the cleaved form of the caspase-3 from western blots of three independent experiments. P value ** < 0.0026