title: LC3-II Tagging and Western Blotting for Monitoring Autophagic Activity in Mammalian Cells.
abstract: The autophagosome-associated protein LC3-II is commonly used as a marker of autophagic activity within cells, but its levels are affected by both formation and degradation of autophagosomes. This can make the significance of altered LC3-II levels ambiguous. Here we describe the method of Bafilomycin A1 blotting, in which the degradation of autophagosomes is prevented in cultured cells, allowing the causes of altered LC3-II levels to be determined.
authors.affiliation: Department of Medical Genetics, Cambridge Institute for Medical Research, University of Cambridge, Cambridge Biomedical Campus, Wellcome Trust/MRC Building, Hills Road, Cambridge, CB2 0XY, UK.
authors.forename: Anne
authors.initials: A
authors.lastname: Streeter
authors.forename: Fiona M
authors.initials: FM
authors.lastname: Menzies
authors.forename: David C
authors.initials: DC
authors.lastname: Rubinsztein
mesh_terms: D001343:Autophagy / Q000187:drug effects; D015153:Blotting, Western / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D018942:Macrolides / Q000494:pharmacology; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D059748:Proteolysis / Q000187:drug effects
chemical_list: C477012:MAP1LC3A protein, human; D018942:Macrolides; D008869:Microtubule-Associated Proteins; C040929:bafilomycin A1