Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

Daniel J. Klionsky, Kotb Abdelmohsen, Akihisa Abe, Md Joynal Abedin, Eeva-Liisa Eskelinen, Juha J. Hulmi, Laura Korhonen, Dan Lindholm, Mark Screen, Vassileios Stratoulias, Susu M. Zughaier

Research output: Contribution to journalArticleScientificpeer-review


More than 2000 authors in alphabetical order.
Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. Along these lines, because of the potential for pleiotropic effects due to blocking autophagy through genetic manipulation, it is imperative to target by gene knockout or RNA interference more than one autophagy-related protein. In addition, some individual Atg proteins, or groups of proteins, are involved in other cellular pathways implying that not all Atg proteins can be used as a specific marker for an autophagic process. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular assays, we hope to encourage technical innovation in the field.
Original languageEnglish
Issue number1
Pages (from-to)1-222
Publication statusPublished - 21 Jan 2016
MoE publication typeA1 Journal article-refereed

Bibliographical note

More than 2000 authors in alphabetical order.

Fields of Science

  • 3111 Biomedicine
  • 1182 Biochemistry, cell and molecular biology
  • Autophagy
  • phagophore
  • amphisome
  • autolysosome
  • lysosome

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