title: Analysis of TLR-Induced Metabolic Changes in Dendritic Cells Using the Seahorse XF(e)96 Extracellular Flux Analyzer.
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.
authors.affiliation: Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Leonard R
authors.initials: LR
authors.lastname: Pelgrom
authors.affiliation: Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Alwin J
authors.initials: AJ
authors.lastname: van der Ham
authors.affiliation: Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands. b.everts@lumc.nl.
authors.forename: Bart
authors.initials: B
authors.lastname: Everts
mesh_terms: D000818:Animals; D003713:Dendritic Cells / Q000378:metabolism*; D004734:Energy Metabolism; D006019:Glycolysis; D057166:High-Throughput Screening Assays; D055442:Metabolome; D055432:Metabolomics / Q000379:methods; D051379:Mice; D008928:Mitochondria / Q000378:metabolism; D010085:Oxidative Phosphorylation; D051193:Toll-Like Receptors / Q000378:metabolism*
chemical_list: D051193:Toll-Like Receptors
keywords: Cellular bioenergetics; Dendritic cells; Extracellular Flux Analyzer; Extracellular acidification; Glycolysis; Metabolism; Oxidative phosphorylation; Oxygen consumption; Respiration; Seahorse Bioscience; Toll-like receptors