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To analyze real-time glycolytic rates, Seahorse XF glycolytic rate assay utilizes both (extracellular acidification rate) ECAR and OCR measurements to evaluate the glycolytic proton efflux rate (glycoPER) of the cells, in which the cells were incubated in glucose-free media followed by the addition of rotenone, antimycin A, and finally 2-deoxyglucose (2-DG, glycolysis inhibitor)

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