Cells were seeded onto 96- or 24-well plates and cultured for 8 hours. The cells were then depleted in culture medium supplemented with 2 or 5% of in-house dialyzed fetal bovine serum (dialyzed against 0.15 M NaCl until glucose is <5 mg/dl; 10,000 molecular weight cutoff) for an additional 18 hours (unless otherwise indicated following drug treatments). Glucose uptake was analyzed using bioluminescent-based Glucose Uptake-Glo Assay (Promega) according to the manufacturer’s instructions. For measuring lactate secretion, cells were replenished with fresh medium after depletion and cultured 36 hours longer. Lactate levels in conditioned medium were measured using a colorimetric-based Lactate Assay Kit (Cell Biolabs). For measuring glutamine consumption, we used the bioluminescent-based Glutamine/Glutamate-Glo Assay (Promega) following the manufacturer’s protocol. Glutamine uptake was terminated by adding 6 mM glutamine to yield a final concentration of 4 mM. Luminescence was measured using a POLARstar Omega luminometer.

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