Guidelines for the use and interpretation of assays for monitoring autophagy (2nd edition)

KLIONSKI, DJ; ABDELMOHSEN, K; ......; IRIBARREN, P; .....

AUTOPHAGY

LANDES BIOSCIENCE

Lugar: Austin, Texas; Año: 2016

1554-8627

Here, we present aset of guidelines for the selection and interpretation of methods for use byinvestigators who aim to examine macroautophagy and related processes, as wellas for reviewers who need to provide realistic and reasonable critiques ofpapers that are focused on these processes. These guidelines are not meant tobe a formulaic set of rules, because the appropriate assays depend in part onthe question being asked and the system being used. In addition, we emphasizethat no individual assay is guaranteed to be the most appropriate one in everysituation, and we strongly recommend the use of multiple assays to monitor autophagy.Along these lines, because of the potential for pleiotropic effects due toblocking autophagy through genetic manipulation it is imperative to delete or knockdown more than one autophagy-related gene. In addition, some individual Atgproteins, or groups of proteins, are involved in other cellular pathways so notall 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 informationcan, or cannot, be obtained from them.