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NCL Method GTA-11

By Stephan Stern, Chris McLeland1, Jamie Rodriguez1

Leidos Biomedical Research, Inc.

Autophagic Dysfunction Assay: Qualitative Analysis of MAP LC3-I to LC3-II Conversion By Western Blot

Listed in Datasets | publication by group NCL Protocols

Version 2.0 - published on 09 Jul 2020 doi:10.17917/WY7Z-MD10 - cite this

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Lysosomal dysfunction is recognized as a potential toxic mechanism for xenobiotics that can result in various pathological states (1).  There is concern that nanoparticles, in particular, may cause lysosomal pathologies, since they are likely to accumulate within lysosomes (2).  Lysosomal dysfunction could potentially result from nanoparticle biopersistance, or inhibition of lysosomal enzymes, such as inhibition of phospholipase resulting in phospholipidosis, or inhibition of lysosomal protein degradation resulting in lysosomal overload (1).  Possibly related to lysosomal dysfunction, nanoparticle exposure has also been shown to cause autophagic dysfunction (3), resulting in accumulation of autophagic vacuoles.  Common methods used to detect autophagic dysfunction include microscopy and protein modification assays, such as microtubule associated protein light chain 3-I (MAP LC3-I) lipidation (4).

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