Cytosolic LC3 Ratio as a Sensitive Index of Macroautophagy in Isolated Rat Hepatocytes and H4-II-E Cells

Citations Over TimeTop 11% of 2007 papers

Abstract

Macroautophagy, an intracellular bulk degradation process in eukaryotes, is sensitive to nutrient supply and deprivation. Microtubule-associated protein 1 light chain 3 (LC3), a mammalian homologue of yeast Atg8, plays an indispensable role in macroautophagy formation and is a suitable marker for this process. Through analysis of the subcellular distribution of LC3, we determined that the cytosolic fraction contained not only a precursor form (LC3-I), but also an apparent active form (LC3-IIs). Both cytosolic LC3-I and LC3-IIs were more responsive to amino acids than those of total homogenate. Moreover, changes in the LC3-IIs/I ratio reflected those in the total proteolytic flux remarkably in both fresh rat hepatocytes and H4-II-E cell lines. Thus, in addition to a sensitive index of macroautophagy, calculating the cytosolic LC3 ratio became an easy and quick quantitative method for monitoring its regulation in hepatocytes and H4-II-E cells.

Related Papers

• → Regulation of selective autophagy: the p62/SQSTM1 paradigm(2017)728 cited
• → Atg4 recycles inappropriately lipidated Atg8 to promote autophagosome biogenesis(2012)235 cited
• → Analysis of Autophagic Activity Using ATG8 Lipidation Assay in Arabidopsis thaliana(2018)8 cited
• → How to reduce the accumulation of autophagic vacuoles in NPC1-deficient neurons: A comparison of two pharmacological strategies(2014)9 cited
• → Autophagy and Vacuole Formation(2017)2 cited