CaltechAUTHORS
Published February 8, 2021 | Version public
Journal Article Metadata-only

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

Creators

  • 1. ROR icon Duke University

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 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.

Additional Information

© 2020 Informa UK Limited, trading as Taylor & Francis Group. Received 14 Jul 2020, Published online: 08 Feb 2021. Dedicated to the memory of our colleagues Caty Casas, Yukiko Kabeya and Beth Levine. In a rapidly expanding and highly dynamic field such as autophagy, it is possible that some authors who should have been included on this manuscript have been missed. D.J.K. extends his apologies to researchers in the field of autophagy who, due to oversight or any other reason, could not be included on this manuscript. Funding: 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. Disclaimer: The views expressed in this article are those of the authors and do not necessarily reflect the official policy or position of the U.S. Food and Drug Administration and the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government. No potential conflict of interest was reported by the corresponding author.

Additional details

Identifiers

Eprint ID
108555
DOI
10.1080/15548627.2020.1797280
Resolver ID
CaltechAUTHORS:20210325-104704700

Related works

Describes
10.1080/15548627.2020.1797280 (DOI)

Funding

NIH
GM131919

Dates

Created
2021-03-30
Created from EPrint's datestamp field
Updated
2023-03-16
Created from EPrint's last_modified field

Caltech Custom Metadata

Caltech groups
Division of Biology and Biological Engineering (BBE)