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Review
. 2007 Nov-Dec;3(6):542-5.
doi: 10.4161/auto.4600. Epub 2007 Jun 19.

How to interpret LC3 immunoblotting

Noboru Mizushima  1 , Tamotsu Yoshimori
Affiliations
Review

How to interpret LC3 immunoblotting

Noboru Mizushima et al. Autophagy. 2007 Nov-Dec.

Abstract

Microtubule-associated protein light chain 3 (LC3) is now widely used to monitor autophagy. One approach is to detect LC3 conversion (LC3-I to LC3-II) by immunoblot analysis because the amount of LC3-II is clearly correlated with the number of autophagosomes. However, LC3-II itself is degraded by autophagy, making interpretation of the results of LC3 immunoblotting problematic. Furthermore, the amount of LC3 at a certain time point does not indicate autophagic flux, and therefore, it is important to measure the amount of LC3-II delivered to lysosomes by comparing LC3-II levels in the presence and absence of lysosomal protease inhibitors. Another problem with this method is that LC3-II tends to be much more sensitive to be detected by immunoblotting than LC3-I. Accordingly, simple comparison of LC3-I and LC3-II, or summation of LC3-I and LC3-II for ratio determinations, may not be appropriate, and rather, the amount of LC3-II can be compared between samples.

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