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Western blot analysis of HEK-293 cells transfected either with empty vector, or native human ATP5A1 (5A1), or cysteine 244 mutant (C244S) human ATP5A1, or cysteine 294 mutant (C294S) human ATP5A1, or the double mutant C244S/C294S human ATP5A1 plasmids for 48 hrs. Cell lysates were incubated with NaHS (100 μM) for an additional 30 min at 37°C. The biotin switch assay using anti-tetraHis antibody was used to differentiate the recombinant ATP5A1 from the endogenous ATP5A1.
