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


Daniel J. Klionsky, University of Michigan, Ann Arbor
Amal Kamal Abdel-Aziz, Ain Shams University
Sara Abdelfatah, Johannes Gutenberg-Universität Mainz
Mahmoud Abdellatif, Medizinische Universität Graz
Asghar Abdoli, Pasteur Institute of Iran
Steffen Abel, Leibniz Institut fur Pflanzenbiochemie
Hagai Abeliovich, Hebrew University of Jerusalem
Marie H. Abildgaard, Kræftens Bekæmpelse
Yakubu Princely Abudu, UiT The Arctic University of Norway
Abraham Acevedo-Arozena, Hospital Universitario de Canarias
Iannis E. Adamopoulos, University of California, Davis
Khosrow Adeli, University of Toronto
Timon E. Adolph, Medizinische Universitat Innsbruck
Annagrazia Adornetto, Università della Calabria
Elma Aflaki, National Eye Institute (NEI)
Galila Agam, Ben-Gurion University of the Negev
Anupam Agarwal, The University of Alabama at Birmingham
Bharat B. Aggarwal, Inflammation Research Center
Maria Agnello, Università degli Studi di Palermo
Patrizia Agostinis, Departement Cellulaire en Moleculaire Geneeskunde
Javed N. Agrewala, Indian Institute of Technology Ropar
Alexander Agrotis, University of Dundee
Patricia V. Aguilar, UT Medical Branch at Galveston
S. Tariq Ahmad, Colby College
Zubair M. Ahmed, University of Maryland School of Medicine
Ulises Ahumada-Castro, Universidad Mayor
Sonja Aits, Institutionen för Experimentell Medicinsk Vetenskap
Shu Aizawa, Nihon University
Yunus Akkoc, Koç Üniversitesi
Tonia Akoumianaki, University of Crete Medical School
Hafize Aysin Akpinar, The Institute of Cancer Research, London
Ahmed M. Al-Abd, Gulf Medical University
Lina Al-Akra, The University of Sydney
Abeer Al-Gharaibeh

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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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