Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1

Daniel Klionsky; Amal Kamal Abdel-Aziz; Sara Abdelfatah; Mahmoud Abdellatif; Asghar Abdoli; Steffen Abel; Hagai Abeliovich; Marie Abildgaard; Yakubu Princely Abudu; Abraham Acevedo-Arozena; Iannis Adamopoulos; Khosrow Adeli; Timon Adolph; Annagrazia Adornetto; Elma Aflaki; Galila Agam; Anupam Agarwal; Bharat Aggarwal; Maria Agnello; Javed Agrewala; Patrizia Agostinis; Alexander Agrotis; Patricia Aguilar; S Tariq Ahmad; Zubair Ahmed; Ulises Ahumada-Castro; Sonja Aits; Shu Aizawa; Yunus Akkoc; Tonia Akoumianaki; Hafize Aysin Akpinar; Ahmed Al-Abd; Lina Al-Akra; Abeer Al-Gharaibeh; Moulay Alaoui-Jamali; Simon Alberti; Elísabet Alcocer-Gómez; Cristiano Alessandri; Muhammad Ali; M Abdul Alim Al-Bari; Saeb Aliwaini; Javad Alizadeh; Eugènia Almacellas; Alexandru Almasan; Alicia Alonso; Guillermo D Alonso; Nihal Altan-Bonnet; Dario C Altieri; Élida M. C Álvarez; Sara Alves; Cristine Alves Da Costa; Mazen Alzaharna; Marialaura Amadio; Consuelo Amantini; Cristina Amaral; Susanna Ambrosio; Amal Amer; Veena Ammanathan; Zhenyi An; Stig Andersen; Shaida Andrabi; Magaiver Andrade-Silva; Allen Andres; Sabrina Angelini; David Ann; Uche Anozie; Mohammad Ansari; Pedro Antas; Adam Antebi; Zuriñe Antón; Tahira Anwar; Lionel Apetoh; Nadezda Apostolova; Toshiyuki Araki; Yasuhiro Araki; Kohei Arasaki; Wagner Araújo; Jun Araya; Catherine Arden; Maria-Angeles Arévalo; Sandro Arguelles; Esperanza Arias; Jyothi Arikkath; Hirokazu Arimoto; Aileen Ariosa; Darius Armstrong-James; Laetitia Arnauné-Pelloquin; Angeles Aroca; Daniela Arroyo; Ivica Arsov; Rubén Artero; Dalia Maria Lucia Asaro; Michael Aschner; Milad Ashrafizadeh; Osnat Ashur-Fabian; Atanas Atanasov; Alicia Au; Patrick Auberger; Holger Auner; RACHEL SIMMONDS; Laure Aurelian

doi:10.1080/15548627.2020.1797280

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Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1

Journal article

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Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1

Daniel Klionsky, Amal Kamal Abdel-Aziz, Sara Abdelfatah, Mahmoud Abdellatif, Asghar Abdoli, Steffen Abel, Hagai Abeliovich, Marie Abildgaard, Yakubu Princely Abudu, Abraham Acevedo-Arozena, …

Autophagy, Vol.17(1), pp.1-382

08/02/2021

DOI: https://doi.org/10.1080/15548627.2020.1797280

PMID: 33634751

Abstract

Autophagosome

Cancer

Flux

LC3

Lysosome

Macroautophagy

Neurodegeneration

Phagophore

Vacuole

Life Sciences

Stress

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field.

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Title: Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1
Creators: Daniel Klionsky - Life Sciences Institute and Department of Molecular, Cellular, and Developmental Biology and Biological Chemistry
Amal Kamal Abdel-Aziz - Department of Pharmacology and Toxicology
Sara Abdelfatah - Department of Pharmaceutical Biology
Mahmoud Abdellatif - Division of Cardiology
Asghar Abdoli - Department of Hepatitis and HIV
Steffen Abel - Department of Molecular Signal Processing
Hagai Abeliovich - Department of Biochemistry and Food Science
Marie Abildgaard - RNA and Autophagy Group
Yakubu Princely Abudu - Department of Medical Biology
Abraham Acevedo-Arozena - CIBERNED and Universidad de La Laguna, ITB
Iannis Adamopoulos - UC Davis Division of Rheumatology, Allergy and Clinical Immunology, School of Medicine
Khosrow Adeli - Department of Biochemistry [University of Toronto]
Timon Adolph - Department of Internal Medicine, Innsbruck Medical University, Innsbruck, Austria
Annagrazia Adornetto - Department of Pharmacy, Health and Nutritional Sciences
Elma Aflaki - Protein Structure and Function Section
Galila Agam - Department of Clinical Biochemistry and Pharmacology
Anupam Agarwal - Division of Nephrology and Birmingham VA Medical Center
Bharat Aggarwal - Inflammation Research Center
Maria Agnello - Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche (STEBICEF)
Javed Agrewala - Indian Institute of Technology
Patrizia Agostinis - Cell Death Research & Therapy (CDRT) Lab, Department of Cellular & Molecular Medicine
Alexander Agrotis - The MRC Protein Phosphorylation and Ubiquitylation Unit [Dundee, UK]
Patricia Aguilar - Department of Pathology [Dallas]
S Tariq Ahmad - Department of Biology
Zubair Ahmed - Departments of Otorhinolaryngology Head & Neck Surgery, Ophthalmology and Visual Science
Ulises Ahumada-Castro - Center for Integrative Biology, and Geroscience Center for Brain Health and Metabolism
Sonja Aits - Department of Experimental Medical Science, Cell Death, Lysosomes and Artificial Intelligence Group
Shu Aizawa - College of Bioresource Sciences, Nihon University
Yunus Akkoc - Department of Molecular Biology and Genetics and KUTTAM Research Center for Translational Medicine
Tonia Akoumianaki - School of Medicine. Laboratory of Clinical Microbiology and Microbial Pathogenesis
Hafize Aysin Akpinar - Cancer Therapeutics Unit
Ahmed Al-Abd - Pharmaceutical Sciences Department, College of Pharmacy
Lina Al-Akra - Department of Pathology and Bosch Institute, University of Sydney
Abeer Al-Gharaibeh - Department of Research
Moulay Alaoui-Jamali - Departments of Medicine and Oncology and Segal Cancer Centre
Simon Alberti - Center for Molecular and Cellular Bioengineering [TU Dresden, Germany]
Elísabet Alcocer-Gómez - Departamento de Psicología Experimental
Cristiano Alessandri - Department of Clinical, Internal, Anesthesiological and Cardiovascular Sciences
Muhammad Ali - Johns Hopkins University School of Medicine [Baltimore]
M Abdul Alim Al-Bari - Department of Pharmacy, Faculty of Science, University of Rajshahi, Rajshahi, Bangladesh
Saeb Aliwaini - Islamic University of Gaza
Javad Alizadeh - University of Manitoba [Winnipeg]
Eugènia Almacellas - University of Barcelona
Alexandru Almasan - Cleveland Clinic
Alicia Alonso - University of the Basque Country [Bizkaia]
Guillermo D Alonso - Instituto de Investigaciones en Ingeniería Genética y Biología Molecular, “Dr. Héctor N. Torres” [Buenos Aires]
Nihal Altan-Bonnet - National Heart, Lung, and Blood Institute [Bethesda]
Dario C Altieri - The Wistar Institute
Élida M. C Álvarez - University of Buenos Aires [Argentina]
Sara Alves - Universidade do Porto
Cristine Alves Da Costa - Excellence Laboratory LabEx DISTALZ
Mazen Alzaharna - Islamic University of Gaza
Marialaura Amadio - University of Pavia
Consuelo Amantini - University of Camerino
Cristina Amaral - Universidade do Porto
Susanna Ambrosio - Telethon Institute of Genetics and Medicine = Istituto Telethon di Genetica e Medicina
Amal Amer - Ohio State University [Columbus]
Veena Ammanathan
Zhenyi An - University of California [San Francisco]
Stig Andersen - Aarhus University [Aarhus]
Shaida Andrabi - University of Alabama at Birmingham [ Birmingham]
Magaiver Andrade-Silva - Federal University of Sao Paulo
Allen Andres - Cedars-Sinai Medical Center
Sabrina Angelini - University of Bologna
David Ann - City of Hope Comprehensive Cancer Center [Duarte]
Uche Anozie - The University of Tennessee [Knoxville]
Mohammad Ansari - Northeastern Ohio Medical University
Pedro Antas - Chronic Diseases Research Center
Adam Antebi - Max planck Institute for Biology of Ageing [Cologne]
Zuriñe Antón
Tahira Anwar
Lionel Apetoh
Nadezda Apostolova
Toshiyuki Araki
Yasuhiro Araki
Kohei Arasaki
Wagner Araújo
Jun Araya
Catherine Arden
Maria-Angeles Arévalo
Sandro Arguelles
Esperanza Arias
Jyothi Arikkath
Hirokazu Arimoto
Aileen Ariosa
Darius Armstrong-James
Laetitia Arnauné-Pelloquin - Centre de Biologie Intégrative
Angeles Aroca
Daniela Arroyo
Ivica Arsov
Rubén Artero
Dalia Maria Lucia Asaro
Michael Aschner
Milad Ashrafizadeh
Osnat Ashur-Fabian
Atanas Atanasov
Alicia Au
Patrick Auberger - Centre méditerranéen de médecine moléculaire
Holger Auner
RACHEL SIMMONDS - University of Surrey, School of Biosciences
Laure Aurelian
Publication Details: Autophagy, Vol.17(1), pp.1-382
Publisher: Taylor & Francis
First online publication date: 08/02/2021
Grant note: This work was supported by the National Institute of General Medical Sciences [GM131919]. Due to space and other limitations, it is not possible to include all other sources of financial support. Grants-in-Aid for Scientific Research: 20K08460, 20K07910, 20K07896, 20K08662, 23K21397, 20K06636, 20K21769, 20H05504, 20K06501, 20K21531, 20J00363, 20H03249, 20H02990, 20H05772, 21K07926, 19K24690, 21H02812 National Eye Institute: R01EY029675 National Heart Lung and Blood Institute: R01HL088256, R01HL141759, R01HL133545 National Institute of Allergy and Infectious Diseases: R01AI136921, R01AI139046 National Institute of Arthritis and Musculoskeletal and Skin Diseases: R01AR061370 National Institute of Diabetes and Digestive and Kidney Diseases: R01DK121545, P30DK020572 National Institute of General Medical Sciences: R35GM122536, R35GM131919, R35GM130331 National Institute on Aging: R56AG063820 BBSRC: BBS/E/T/000PR9814, BBS/E/B/000C0434, BBS/E/F/000PR10355, BB/T006102/1, BBS/E/B/000C0413, BBS/E/F/00044500, BB/R008167/1, BB/R019258/1, BBS/E/T/000PR9819 FLF: MR/S032304/1 MRC: MC_UU_00018/2, MR/T002220/1, MR/M00869X/2, G0501003, MR/N022696/1, MC_U105184308, MR/R009732/1, MR/S009426/1, MC_EX_G0800785, MR/R025096/1
Identifiers: 99550921102346; WOS:000636121800001
Academic Unit: School of Biosciences
Language: English
Resource Type: Journal article
SDG (Scopus 2023): SDG 3

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1

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