Fig. 3: SLC25A15 deletion inhibits mitochondrial activity and UC in the mut-ER BC cells.

A SLC25A15 gene was edited using CRISPR/cas9 and clones of WT-ER or mut-ER cells were generated, then cells were lysed and immunoblotted with SLC25A15. β-actin served as a control and band intensity was quantified using ImageJ. B Mitochondrial activity of WT-ER and Y537S-ER or C WT-ER and D538G-ER MCF-7 cells was studied by monitoring OCR using Seahorse. Mitochondrial respiration was measured using cell mito stress test kit under basal conditions followed by the sequential addition of oligomycin (2 mM), FCCP (0.5 mM), rotenone (0.5 mM). Both basal OCR and coupling efficiency were quantified. OCR values were normalized to cell numbers by DAPI staining. To exclude non-mitochondrial oxygen consumption from the analysis, the third OCR value recorded after injection of the rotenone/antimycin A mixture was subtracted from each OCR value. The figures depict a representative graph output of at least three independent experiments. D Urea concentration was measured in the media of the mutated-ER cells (control and knockouts). Absorbance was measured at 570 nm. Urea concentration was normalized to protein concentration. Each bar represents the mean ± S.D. *P < 0.05, **P < 0.01; ***P < 0.001. Con Control, SLC-KO: SLC25A15 knockout.