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Methods Mol Biol. 2018;1713:69-75. doi: 10.1007/978-1-4939-7507-5_6.

Evaluating the Efficacy of GLUT Inhibitors Using a Seahorse Extracellular Flux Analyzer.

Author information

1
Department of Hematology and Medical Oncology, Winship Cancer Institute, School of Medicine, Emory University, Atlanta, GA, 30322, USA.
2
Department of Pediatrics, Washington University School of Medicine, St. Louis, MO, 63110, USA.
3
Department of Hematology and Medical Oncology, School of Medicine, Emory University, 1365C Clifton Rd. NE, Atlanta, GA, 30322, USA. mala.shan@emory.edu.

Abstract

Glucose is metabolized through anaerobic glycolysis and aerobic oxidative phosphorylation (OXPHOS). Perturbing glucose uptake and its subsequent metabolism can alter both glycolytic and OXPHOS pathways and consequently lactate and/or oxygen consumption. Production and secretion of lactate, as a consequence of glycolysis, leads to acidification of the extracellular medium. Molecular oxygen is the final electron acceptor in the electron transport chain, facilitating oxidative phosphorylation of ADP to ATP. The alterations in extracellular acidification and/or oxygen consumption can thus be used as indirect readouts of glucose metabolism and assessing the impact of inhibiting glucose transport through specific glucose transporters (GLUTs). The Seahorse bioenergetics analyzer can measure both the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR). The proposed methodology affords a robust, high-throughput method to screen for GLUT inhibition in cells engineered to express specific GLUTs, providing live cell read-outs upon GLUT inhibition.

KEYWORDS:

ECAR; GLUT inhibitor; Glycolysis; Oxidative phosphorylation; Seahorse XF analyzer

PMID:
29218518
DOI:
10.1007/978-1-4939-7507-5_6
[Indexed for MEDLINE]

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