Measurement of respiration of living cells with full access to all metabolic substrates was performed using a Seahorse XF24 analyzer (20). Respiration (OCR) was measured under basal conditions in the presence of the mitochondrial inhibitor oligomycin (1.2 μM), the mitochondrial uncoupling compound FCCP (5 μM), and the respiratory chain inhibitor rotenone (1 μM). Glycolytic activities were measured simultaneously using the same instrument based on the ECAR. Total cellular ATP levels were assessed using an ATP bioluminescent somatic cell assay kit (Sigma-Aldrich, St. Louis, MO), following the instructions provided by the manufacturer. To measure cellular ROS levels, cells were loaded with 2′-7′-dichlorofluorescein diacetate (5 μM) at 37°C for 15 min. ROS (H2O2) levels were then determined by FACS.

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