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. 2021 Sep 30;12(10):893. doi: 10.1038/s41419-021-04190-w

Fig. 4. OXPHOS inhibitors potentiate the anticancer efficacy of alisertib.

Fig. 4

A Growth inhibition assay in MDA-MB-231 cells treated with alisertib alone (blue lines) or in combination (red lines) with mitochondrial inhibitors, including metformin (5 mM), sodium azide (1 mM), oligomycin (1 nM), or FCCP (10 mM). The relative absorbance was normalized and vehicle-treated cells were set as 100%. B–G Colony-forming assay in the MDA-MB-231 (B), 4T1 (D), HeLa (F) cells treated with 100 nM alisertib, 5 mM metformin, or combination (A + M). Scale bar, 500 μm. H MDA-MB-231 cells in 3D culture. Cells were grown in Matrigel with 100 nM alisertib, 5 mM metformin, or combination treatment. Images were taken on day 5. Scale bar, 500 µm. I, J Quantification of the diameters of cell clusters (I) and the percentage of cell clusters in diameter exceeded 30 μm (J), n = 3. ns not significant; ****p < 0.0001. One-way ANOVA. K Annexin V staining for apoptotic cells was performed in MDA-MB-231 cells after 24 h of treatment with 100 nM alisertib (ALS), 5 mM metformin (Metf), or in combination. Viable cells are in the lower left quadrant, and early apoptotic and late apoptotic cells are in the lower right and upper right quadrants, respectively.