Journal
BIOESSAYS
Volume 42, Issue 11, Pages -Publisher
WILEY
DOI: 10.1002/bies.202000122
Keywords
autolysosome; autophagosome; autophagy; autophagy-flux; gamma-aminobutyric acid receptor-associated protein; microtubule-associated proteins 1A; 1B light chain 3B
Categories
Funding
- Japan Society for the Promotion of Science [19H05706, 18H02611]
- Takeda Science Foundation
- High Technology Research Center Grant, Strategic Research Foundation at Private Universities
- Grants-in-Aid for Scientific Research [19H05706, 18H02611] Funding Source: KAKEN
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Macroautophagy is a major degradation mechanism of cell components via the lysosome. Macroautophagy greatly contributes to not only cell homeostasis but also the prevention of various diseases. Because macroautophagy proceeds through multi-step reactions, researchers often face a persistent question of how macroautophagic activity can be measured correctly. To make a straightforward determination of macroautophagic activity, diverse monitoring assays have been developed. Direct measurement of lysosome-dependent degradation of radioisotopically labeled cell proteins has long been applied. Meanwhile, indirect monitoring procedures have been developed. In these assays, autophagosome marker proteins, microtubule-associated proteins 1A/1B light chain 3B-II (LC3B-II) and gamma-aminobutyric acid receptor-associated protein-II (GABARAP-II) have been analyzed and the validity of the assays strongly depends on appropriate assessment of the fluctuation of LC3-II and/or GABARAP-II levels in the presence or absence of lysosomal inhibitors. This article describes these monitoring methods, paying special attention to the principles and characteristics of each procedure.
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