title: Measuring cardiac autophagic flux in vitro and in vivo.
abstract: Autophagy is a lysosomal-dependent catabolic pathway that recycles various cytoplasmic-borne components, such as organelles and proteins, through the lysosomes. This process creates energy and biomolecules that are used to maintain homeostasis and to serve as an energy source under conditions of acute stress. Autophagic flux is a measure of efficiency or throughput of the pathway. Here, we describe a method for determining autophagic flux in vitro and in vivo using the autophagosomal/lysosomal fusion inhibitors chloroquine or bafilomycin A1 and then probing for the autophagosomal marker LC3-II via Western Blot.
authors.affiliation: Donald P. Shiley BioScience Center, San Diego State University, 5500 Campanile Dr., San Diego, CA, 92182-4650, USA.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Gurney
authors.forename: Chengqun
authors.initials: C
authors.lastname: Huang
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Ramil
authors.forename: Nandini
authors.initials: N
authors.lastname: Ravindran
authors.forename: Allen M
authors.initials: AM
authors.lastname: Andres
authors.forename: Jon
authors.initials: J
authors.lastname: Sin
authors.forename: Phyllis-Jean
authors.initials: PJ
authors.lastname: Linton
authors.forename: Roberta A
authors.initials: RA
authors.lastname: Gottlieb
mesh_terms: D000818:Animals; D001343:Autophagy / Q000187:drug effects; D015153:Blotting, Western / Q000379:methods*; D002738:Chloroquine / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology; D066298:In Vitro Techniques; D018942:Macrolides / Q000494:pharmacology; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009206:Myocardium / Q000473:pathology*
chemical_list: D004791:Enzyme Inhibitors; D018942:Macrolides; C480860:Map1lc3b protein, mouse; D008869:Microtubule-Associated Proteins; D002738:Chloroquine; C040929:bafilomycin A1