Fig. 8

The regulation of the stability of MCL-1, cyclin B1, and other pro- or anti-apoptotic proteins in HeLa and MCF-7 cells. a Western blots of cell extracts prepared from HeLa cells after treatment with 0.5% DMSO, 50 μM M2I-1, 60 ng/ml nocodazole, and 60 ng/ml nocodazole + 50 μM M2I-1 respectively and probed with a rabbit polyclonal anti-cyclin B1 antibody (H-433) antibody (sc-7520); a mouse monoclonal anti-actin (AC-15) antibody (ab6276) was used as the loading control. b–f Western blot results showing the expression levels of the pro-apoptotic proteins BIM, BID, PUMA, NOXA, and MCL-1s in HeLa cells. Cell extracts were prepared from HeLa cells after treatment with nocodazole alone or nocodazole + M2I-1 as described before. Actin bands were used as the loading controls. MCF-7 cells were treated using the same conditions as described in Fig. 2e apart from the determination of the chromosomal morphologies, which was highlighted by staining with 0.2 μ M SiR-DNA. Digital images were taken after 20 h incubation for quantification of the mitotic (g), apoptotic (h), and slippage indices (i). n: The number of the parallel experiments and the total cell numbers used for quantification. P value * < 0.026, 0.029, and 0.036 respectively