title: Fluorometric Methods for Detection of Mitochondrial Membrane Depolarization Induced by CD95 Activation.
abstract: Mitochondria are key organelles implicated in energy supply and apoptosis. Therefore, tracking mitochondria and measuring their membrane potential is of crucial interest to monitor the CD95-mediated apoptotic signal. In this chapter, we report how we evaluate the drop of the mitochondrial transmembrane potential in leukemic cells and adherent triple negative breast cancer cells exposed to cytotoxic CD95L. We describe a simple, robust, and well-established protocol using classical fluorescent probes, DIOC6(3) and TMRM. Living cells are loaded with these cationic dyes, which accumulate in mitochondria. After CD95 activation, organelle depolarization is assessed using flow cytometry.
authors.affiliation: Galapagos SASU, 102 Avenue Gaston Roussel, 93230, Romainville, France.
authors.forename: Kenji F
authors.initials: KF
authors.lastname: Shoji
authors.affiliation: Centre Eugène Marquis, rue Bataille Flandres Dunkerque, 35042, Rennes, France. laure.debure@univ-rennes1.fr.|INSERM ERL440-OSS, Equipe Labellisée Ligue Contre Le Cancer, Rennes, France. laure.debure@univ-rennes1.fr.|Université de Rennes-1, 2 av. du Prof. Léon Bernard, 35043, Rennes, France. laure.debure@univ-rennes1.fr.
authors.forename: Laure
authors.initials: L
authors.lastname: Debure
mesh_terms: D017209:Apoptosis; D053222:Fas Ligand Protein / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods; D005470:Fluorometry / Q000379:methods*; D053078:Membrane Potential, Mitochondrial; D008928:Mitochondria / Q000378:metabolism; D051336:Mitochondrial Membranes / Q000378:metabolism*; D011485:Protein Binding; D012235:Rhodamines / Q000378:metabolism; D019014:fas Receptor / Q000378:metabolism*
chemical_list: D053222:Fas Ligand Protein; D012235:Rhodamines; D019014:fas Receptor; C401833:tetramethylrhodamine methyl ester
keywords: Apoptosis; CD95; CD95L; DIOC6(3); Flow cytometry; Mitochondrial membrane potential; TMRM