Cited by 2,597Autophagy in Disease and TherapyCalcium signaling and nucleotide metabolismCellular transport and secretion

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How to Interpret LC3 Immunoblotting

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Mizushima, Noboru, Yoshimori, Tamotsu·2007·Autophagy

DOI 10.4161/auto.4600

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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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LC3, GABARAP and GATE16 localize to autophagosomal membrane depending on form-II formation
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Authors: Mizushima, N., & Yoshimori, T.
Affiliations: Tokyo Medical and Dental University (JP)
ORCID: Mizushima, Noboru
Yoshimori, Tamotsu
Venue: Autophagy
Publisher: Informa UK Limited
Published: 2007-11-26
Volume: 3
Issue: 6
Pages: 542-545
ISSN: 1554-8627 (ISSN-L)
1554-8635
DOI: 10.4161/auto.4600
OpenAlex: W1963521777
Semantic Scholar: ab5143ed104a787ff2e14d915df9a12ce5f5871a
Cited by: 2,595
References: 21
Citations by year: 2026: 62 · 2025: 114 · 2024: 152 · 2023: 141 · 2022: 176 · 2021: 164 · 2020: 173 · 2019: 155 · 2018: 177 · 2017: 162 · 2016: 167 · 2015: 179 · 2014: 137 · 2013: 165 · 2012: 120
FWCI: 52.76
Type: journal article
Language: en
Open access: hybrid
doi.org
Licence: cc-by-nc
Locations: Autophagy (publishedVersion, cc-by-nc)
PubMed (publishedVersion)
Funders: Kato Memorial Bioscience Foundation
Toray Science Foundation
Japan Society for the Promotion of Science
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Autophagy in Disease and Therapy
Life Sciences › Biochemistry, Genetics and Molecular Biology › Physiology › Calcium signaling and nucleotide metabolism
Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
Keywords: LC3, autophagic flux
MeSH terms: Animals, Autophagy, Humans, Lysosomes, Microtubule-Associated Proteins, Phosphatidylethanolamines, Sensitivity and Specificity, Immunoblotting, PC12 Cells, Cell Line, Tumor, Adaptor Proteins, Signal Transducing, Rats

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