The assay medium was the base medium (or KHB ) supplemented with 5.5 mM glucose and 50 µM carnitine (required to transport long chain fatty acid into the mitochondria). Fatty acids tested include long chain fatty acid palmitate, medium chain fatty acid octanoate, and short chain fatty acid butyrate. They were titrated for concentrations stimulating maximal OCR response. The working concentration of palmitate conjugated with BSA was 150 µM, and octanoate 1 mM, which were also above saturation.
It is critical that the above assay conditions are strictly adhered to. Any variation in the assay medium composition may result in different interpretations and insights into cellular metabolic network. Saturating substrate concentrations and optimal compound concentrations were determined by performing titration experiments as described in Materials S4 and S5 inFile S1 . The effect of assay conditions on the interpretation of experimental results will be described elsewhere.
It is critical that the above assay conditions are strictly adhered to. Any variation in the assay medium composition may result in different interpretations and insights into cellular metabolic network. Saturating substrate concentrations and optimal compound concentrations were determined by performing titration experiments as described in Materials S4 and S5 in
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