title: Measurement of Autophagy by Flow Cytometry.
abstract: Autophagy activation is characterized by the accumulation of double-membrane autophagic vesicles (autophagosomes) in the cytoplasm. The mere presence of autophagosomes in the cytoplasm does not necessarily indicate an increased level of autophagy, since the blockade of any step downstream of autophagosome formation increases the number of autophagosomes. Therefore, quantitative methods for the detection of cytoplasmic protein turnover should be employed in addition to autophagosome monitoring, to verify increased levels of autophagy. At the present, multiple methods are available for the quantification of autophagy and the identification of autophagosomes. Here, we detail the in vitro methods currently available to detect autophagic cell death by flow cytometry analysis.
authors.affiliation: Department of Biochemistry, Biophysics and General Pathology, Second University of Naples, Via L. De Crecchio, 7, Naples, 80138, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Zappavigna
authors.affiliation: Department of Biochemistry, Biophysics and General Pathology, Second University of Naples, Via L. De Crecchio, 7, Naples, 80138, Italy.
authors.forename: Angela
authors.initials: A
authors.lastname: Lombardi
authors.affiliation: Department of Biochemistry, Biophysics and General Pathology, Second University of Naples, Via L. De Crecchio, 7, Naples, 80138, Italy.
authors.forename: Gabriella
authors.initials: G
authors.lastname: Misso
authors.affiliation: Department of Biochemistry, Biophysics and General Pathology, Second University of Naples, Via L. De Crecchio, 7, Naples, 80138, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Grimaldi
authors.affiliation: Department of Biochemistry, Biophysics and General Pathology, Second University of Naples, Via L. De Crecchio, 7, Naples, 80138, Italy.
authors.forename: Michele
authors.initials: M
authors.lastname: Caraglia
mesh_terms: D000071182:Autophagosomes / Q000378:metabolism; D001343:Autophagy; D015415:Biomarkers; D005434:Flow Cytometry / Q000379:methods; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D008247:Lysosomes / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D057054:Molecular Imaging; D013194:Staining and Labeling; D057188:Workflow
chemical_list: D015415:Biomarkers; D008869:Microtubule-Associated Proteins
keywords: Autophagosomes; Cell death; Flow cytometry; LC3; Lysosomes