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LC3 and Autophagy.

Isei Tanida¹, Takashi Ueno, Eiki Kominami

¹Department of Biochemistry, Juntendo University School of Medicine, Bunkyo-ku, Tokyo, Japan.

Methods in Molecular Biology (Clifton, N.J.)

|April 22, 2008

Summary

This summary is machine-generated.

Microtubule-associated protein 1A/1B-light chain 3 (LC3) is a key marker for monitoring autophagy. Assays for LC3-II via immunoblotting, immunoprecipitation, and immunofluorescence are described for studying autophagic activity.

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Area of Science:

Cell Biology
Molecular Biology

Background:

Microtubule-associated protein 1A/1B-light chain 3 (LC3) is a ubiquitous protein in mammalian cells.
LC3 undergoes lipidation to form LC3-II during autophagy, a process essential for autophagosome formation and cargo degradation.
Lysosomal turnover of LC3-II serves as a reliable indicator of autophagic activity.

Purpose of the Study:

To describe basic protocols for assaying endogenous LC3-II.
To provide methods for monitoring autophagy and related processes, including autophagic cell death.

Main Methods:

Immunoblotting to detect LC3-II.
Immunoprecipitation to isolate and detect LC3-II.
Immunofluorescence to visualize LC3-II localization.

Main Results:

Established protocols for detecting endogenous LC3-II using immunoblotting, immunoprecipitation, and immunofluorescence.
Demonstrated the utility of LC3-II detection for monitoring starvation-induced autophagic activity.

Conclusions:

LC3-II detection is a robust method for assessing autophagy.
The described protocols facilitate the study of autophagy and autophagic cell death.