Format

Send to

Choose Destination
Methods. 2015 Mar;75:13-8. doi: 10.1016/j.ymeth.2014.11.021. Epub 2014 Dec 5.

LC3- and p62-based biochemical methods for the analysis of autophagy progression in mammalian cells.

Author information

1
Department of Biochemistry and Molecular Biology, Graduate School and Faculty of Medicine, The University of Tokyo, Tokyo 113-0033, Japan; Department of Physiology and Cell Biology, Tokyo Medical and Dental University, Tokyo 113-8519, Japan.
2
Department of Biochemistry and Molecular Biology, Graduate School and Faculty of Medicine, The University of Tokyo, Tokyo 113-0033, Japan. Electronic address: nmizu@m.u-tokyo.ac.jp.

Abstract

Autophagy is an intracellular degradation system that delivers cytoplasmic materials to the lysosome or vacuole. This system plays a crucial role in various physiological and pathological processes in living organisms ranging from yeast to mammals. Thus, an accurate and reliable measure of autophagic activity is necessary. However, autophagy involves dynamic and complicated processes that make it difficult to analyze. The term "autophagic flux" is used to denote overall autophagic degradation (i.e., delivery of autophagic cargo to the lysosome) rather than autophagosome formation. Immunoblot analysis of LC3 and p62/SQSTM1, among other proteins, has been widely used to monitor autophagic flux. Here, we describe basic protocols to measure the levels of endogenous LC3 and p62 by immunoblotting in cultured mammalian cells.

KEYWORDS:

Autophagic flux; Autophagy; Immunoblotting; LC3; SQSTM1; p62

PMID:
25484342
DOI:
10.1016/j.ymeth.2014.11.021
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center