Analysis of TLR-Induced Metabolic Changes in Dendritic Cells Using the Seahorse XF(e)96 Extracellular Flux Analyzer

Methods in Molecular Biology
Leonard R PelgromBart Everts

Abstract

Engagement of Toll-like receptors (TLRs) on dendritic cells (DCs) triggers the expression of a large set of genes involved in DC activation and maturation, which allow them to act efficiently as antigen-presenting cells. Recently, it has become clear that TLR signalling in DCs also results in dramatic metabolic changes that are integral to their changed biology. Here, we describe a detailed protocol on how DC metabolism can be studied after TLR stimulation using the 96-well format Extracellular Flux (XF(e)96) Analyzer from Seahorse Bioscience, a machine that allows one to simultaneously assess rates of oxidative phosphorylation and glycolysis in real-time, in live cells and in a high-throughput manner.

Citations

Nov 21, 2018·Frontiers in Immunology·Farhan BasitI Jolanda M de Vries
May 10, 2019·Frontiers in Immunology·Qun ZengHerbert Schwarz
Apr 17, 2021·Nature Reviews. Immunology·Kelsey VossJeffrey C Rathmell
Oct 8, 2020·Cell Metabolism·Maxim N Artyomov, Jan Van den Bossche

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