Journal
AUTOPHAGY
Volume 4, Issue 2, Pages 151-175Publisher
TAYLOR & FRANCIS INC
DOI: 10.4161/auto.5338
Keywords
autolysosome; autophagosome; flux; lysosome; phagophore; stress; vacuole
Categories
Funding
- BBSRC [BB/E01030X/1] Funding Source: UKRI
- MRC [G0600194, G0601133] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/E01030X/1] Funding Source: researchfish
- Medical Research Council [G0601133, G0600194] Funding Source: researchfish
- Division Of Integrative Organismal Systems
- Direct For Biological Sciences [0818802] Funding Source: National Science Foundation
- Biotechnology and Biological Sciences Research Council [BB/E01030X/1] Funding Source: Medline
- Medical Research Council [G0600194(77639), G0601133, G0600194] Funding Source: Medline
- NCI NIH HHS [R01 CA023378-29, R01 CA023378] Funding Source: Medline
- NEI NIH HHS [R01 EY013520] Funding Source: Medline
- NIAID NIH HHS [R01 AI073099, P01 AI056097, R01 AI079065] Funding Source: Medline
- NIAMS NIH HHS [R01 AR055255-02, R01 AR055255] Funding Source: Medline
- NIA NIH HHS [R01 AG026389, R01 AG026389-01A2] Funding Source: Medline
- NIGMS NIH HHS [R01 GM062509, R01 GM075061-03, GM53396, R01 GM075061, R01 GM053396] Funding Source: Medline
- Telethon [TCR04004] Funding Source: Medline
- Wellcome Trust [064354] Funding Source: Medline
Ask authors/readers for more resources
Research in autophagy continues to accelerate,1 and as a result many new scientists are entering the field. Accordingly, it is important to establish a standard set of criteria for monitoring macroautophagy in different organisms. Recent reviews have described the range of assays that have been used for this purpose.(2,3) There are many useful and convenient methods that can be used to monitor macroautophagy in yeast, but relatively few in other model systems, and there is much confusion regarding acceptable methods to measure macroautophagy in higher eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers of autophagosomes versus those that measure flux through the autophagy pathway; thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from fully functional autophagy that includes delivery to, and degradation within, lysosomes (in most higher eukaryotes) or the vacuole (in plants and fungi). Here, we present a set of guidelines for the selection and interpretation of the methods that can be used by investigators who are attempting to examine macroautophagy and related processes, as well as by reviewers who need to provide realistic and reasonable critiques of papers that investigate these processes. This set of guidelines is 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 verify an autophagic response.
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