Autophagy pp 535-540 | Cite as

Measuring Nonselective and Selective Autophagy in the Liver

  • Takashi UenoEmail author
  • Masaaki KomatsuEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1880)


Administration of leupeptin, a specific inhibitor of lysosomal cysteine proteinases, to starved rats or mice inhibits autolysosomal protein degradation and results in accumulation of autolysosomes in their livers. Immunoblotting of liver homogenates to examine autophagic flux in vivo reveals elevated levels of the selective autophagy substrate p62 and the autophagosomal membrane protein LC3-II in the livers of leupeptin-treated animals. Percoll density gradient centrifugation can be used to isolate autolysosomes from the livers of untreated and leupeptin-treated animals. Moreover, autolysosomes can be examined for the presence of sequestered cytoplasmic proteins as well as degradation intermediates.

Key words

Liver Leupeptin Autolysosome Percoll Selective autophagy Nonselective autophagy 



M.K. is supported by Grant-in-Aid for Scientific Research on Innovative Areas (25111006 and 25111001, to M.K) and Japan Society for the Promotion of Science (an A3 foresight program). T.U. is supported by High Technology Research Center Grant, Strategic Research Foundation at Private Universities. T.U. and M.K. are supported by Grant-in-Aid for Scientific Research on Priority Areas (18076005) and the Takeda Science Foundation.


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Laboratory of Proteomics and Biomolecular Science, Research Support CenterJuntendo University Graduate School of MedicineTokyoJapan
  2. 2.Department of PhysiologyJuntendo University Graduate School of MedicineTokyoJapan

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