Fig. 5: The combination of alisertib and metformin prolongs mitosis and increases mitotic cell death.

A Cell cycle analysis of MDA-MB-231 cells treated with the indicated drugs for 24 h. B Quantification of 4n cells treated with the indicated drugs for 24 h in Fig. 5A. Statistical analysis was performed by one-way ANOVA. ns not significant; *p < 0.05; ***p < 0.001. C The representative images of cell division were recorded by time-lapse microscopy. The duration from NEBD to cytokinesis and until death or slippage was monitored. Scale bar, 5 µm. The duration from NEBD to cytokinesis or until DiM were measured and plotted as bar graph. The length indicates the duration of mitosis and its color represents the cell fate. D–I Mitosis duration and the fates of cell division recorded by time-lapse microscopy in MDA-MB-231 (D), 4T1 (F), and HeLa (H) cells. Cells were grown in complete culture medium with 100 nM alisertib, 5 mM metformin, combination (Alisertib + Metformin), or combination supplemented with 5 mM pyruvate (ALS + Metf + PYR). Duration of mitosis for MDA-MB-231 (E), 4T1 (G), and HeLa (I) cells treated with 100 nM alisertib (ALS), 5 mM metformin (Metf), combination (A + M), or combination supplemented with 5 mM pyruvate (A + M + P). J pH-corrected PercevalHR ratio measured by flow cytometry in MDA-MB-231 cells synchronized at mitosis, and treated with DMSO, 5 mM metformin (Metf), 100 nM alisertib, combination (A + M), or combination with 5 mM pyruvate (A + M + P). Analysis by one-way ANOVA. ns not significant; *p < 0.05; ****p < 0.0001. Mean ± SD, n = 3.