Figure EV2. Respiration rates of investigated cells under different metabolic settings.

1. Determination of OXPHOS activity by monitoring oxygen consumption rates (OCR) and extracellular acidification rates (ECAR). After monitoring basal respiration, oligomycin (1 µM), uncoupler FCCP (trifluoromethoxy carbonylcyanide phenylhydrazone; 0.5 µM), rotenone, and antimycin A (Rot, 0.5 µM; AA, 0.5 µM) were added at the indicated times to determine CV‐linked respiration, maximum respiration, and OXPHOS‐specific oxygen consumption, respectively. OCR and ECAR were determined with an automatic flux analyzer (96XF, Seahorse/Agilent). ˜30,000 cells per well were seeded the day before measurement. Cells were supplied with high glucose (25 mM, HGlc), low glucose (5.6 mM), or galactose (10 mM, Gal) for 3 weeks. OCRs were in addition normalized on the relative mitochondrial mass. The error bars denote SEM.
2. Basal, CV‐linked, and maximal respiration rates in respiratory cells with different IF1 levels. Three technical replicates from same clones, each technical replicate with 5 parallel measurements. Spare = reserve capacity, difference between basal and maximal respiration; Proton leak = respiration that is not linked to CV (proton leakage as a result of incomplete coupling); Non‐mito = Respiration that is not blocked upon addition of electron transport chain inhibitors. The error bars denote SEM.