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

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Glucose Transport

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1713))

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.

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Author information

Authors and Affiliations

  1. Department of Hematology and Medical Oncology, Winship Cancer Institute, School of Medicine, Emory University, Atlanta, GA, 30322, USA

    Changyong Wei

  2. Department of Pediatrics, Washington University School of Medicine, St. Louis, MO, 63110, USA

    Monique Heitmeier & Paul W. Hruz

  3. Department of Hematology and Medical Oncology, School of Medicine, Emory University, 1365C Clifton Rd. NE, Atlanta, GA, 30322, USA

    Mala Shanmugam

Authors
  1. Changyong Wei
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  2. Monique Heitmeier
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  3. Paul W. Hruz
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  4. Mala Shanmugam
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Corresponding author

Correspondence to Mala Shanmugam .

Editor information

Editors and Affiliations

  1. Department of Experimental Medical Science, Lund University, Lund, Sweden

    Karin Lindkvist-Petersson

  2. Department of Experimental Medical Science, Lund University, Lund, Sweden

    Jesper S Hansen

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Wei, C., Heitmeier, M., Hruz, P.W., Shanmugam, M. (2018). Evaluating the Efficacy of GLUT Inhibitors Using a Seahorse Extracellular Flux Analyzer. In: Lindkvist-Petersson, K., Hansen, J. (eds) Glucose Transport. Methods in Molecular Biology, vol 1713. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7507-5_6

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  • DOI: https://doi.org/10.1007/978-1-4939-7507-5_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7506-8

  • Online ISBN: 978-1-4939-7507-5

  • eBook Packages: Springer Protocols

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