title: Assays for Monitoring Macroautophagy Activity in T cells.
abstract: Autophagy is an essential catabolic process that regulates a diverse array of functions by targeting cellular components for degradation by lysosomes. Studies in mammalian cells have shown that the regulation of autophagy is highly complex and optimization of experimental approaches to analyze this process needs to be developed for each model studied. This chapter provides an overview of two of the most commonly used ways to monitor autophagy activity in T cell. It involves description of common techniques, namely Western blot and cell immunostaining, giving specific recommendations for working with T cells and monitoring macroautophagy. We also discuss the analysis required for correct interpretation of the results and quantification of macroautophagy activity.
authors.affiliation: Department of Pathology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY, 10461, USA. yair.botbol@einstein.yu.edu.
authors.forename: Yair
authors.initials: Y
authors.lastname: Botbol
authors.affiliation: Department of Pathology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY, 10461, USA. fernando.macian@einstein.yu.edu.
authors.forename: Fernando
authors.initials: F
authors.lastname: Macian
mesh_terms: D000818:Animals; D001343:Autophagy / Q000502:physiology; D002469:Cell Separation / Q000379:methods; D005455:Fluorescent Antibody Technique; D008213:Lymphocyte Activation / Q000276:immunology; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D016176:T-Lymphocyte Subsets / Q000378:metabolism; D013601:T-Lymphocytes / Q000378:metabolism*
chemical_list: C480860:Map1lc3b protein, mouse; D008869:Microtubule-Associated Proteins
keywords: LC3; Lysosome; Macroautophagy; T cell