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Methods Mol Biol. 2018;1846:325-334. doi: 10.1007/978-1-4939-8712-2_22.

Metabolic Analysis of Lymphatic Endothelial Cells.

Author information

1
Department of Internal Medicine, Yale Cardiovascular Research Center, Yale University School of Medicine, New Haven, CT, USA.
2
Section of Endocrinology, Department of Internal Medicine, Yale University School of Medicine, New Haven, CT, USA.
3
Department of Internal Medicine, Yale Cardiovascular Research Center, Yale University School of Medicine, New Haven, CT, USA. michael.simons@yale.edu.
4
Department of Cell Biology, Yale University School of Medicine, New Haven, CT, USA. michael.simons@yale.edu.

Abstract

Metabolism is pivotal for formation of the lymphatic vasculature. Understanding metabolism in lymphatic endothelial cells (LECs) requires quantitative characterization of specific metabolic pathways. Here we describe methods for using radioactive tracers to assess flux rates of glycolysis, fatty acid β-oxidation, glucose oxidation, and glutamine oxidation. We also provide a detailed method for utilizing mass spectrometry (MS) to measure glycolytic intermediates and ATP.

KEYWORDS:

Cellular metabolism; Fatty acid β-oxidation; Glucose oxidation; Glutamine oxidation; Glycolysis; Lymphatic endothelial cells; Mass spectrometry

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