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

Article


Klionsky, D., Roberts, H. and et al. 2021. Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition). Autophagy. 17 (1), pp. 1-382. https://doi.org/10.1080/15548627.2020.1797280
TypeArticle
TitleGuidelines for the use and interpretation of assays for monitoring autophagy (4th edition)
AuthorsKlionsky, D., Roberts, H. and et al.
Abstract

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 non-canonical 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.

KeywordsAutophagosome; cancer; flux; LC3; lysosome; macroautophagy; neurodegeneration; phagophore; stress; vacuole
PublisherTaylor & Francis (Routledge)
JournalAutophagy
ISSN1554-8627
Electronic1554-8635
Publication dates
Print02 Jan 2021
Online08 Feb 2021
Publication process dates
Deposited09 Dec 2020
Accepted19 Oct 2020
Output statusPublished
Accepted author manuscript
Copyright Statement

This is an Accepted Manuscript of an article published by Taylor & Francis in Autophagy on 02/01/2021, available online: http://www.tandfonline.com/10.1080/15548627.2020.1797280

Digital Object Identifier (DOI)https://doi.org/10.1080/15548627.2020.1797280
PubMed ID33634751
Web of Science identifierWOS:000636121800001
LanguageEnglish
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