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Autophagy pp 295-303 | Cite as

Imaging Noncanonical Autophagy and LC3-Associated Phagocytosis in Cultured Cells

  • Elise Jacquin
  • Katherine Fletcher
  • Oliver FloreyEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1880)

Abstract

Monitoring of ATG8 proteins by western blotting and immunofluorescence microscopy are the most common methods to monitor the autophagy pathway. However, it has recently been shown that ATG8 proteins can be lipidated to non-autophagosome, single-membrane compartments through a noncanonical autophagy pathway. This is commonly found to occur during macro-endocytic processes such as phagocytosis, where it has been termed LC3-associated phagocytosis, and upon lysosomotropic drug treatment. Therefore, care is required when interpreting data based on ATG8 in order to conclude whether a signal relates to the canonical or noncanonical pathway. Here we provide methods to monitor noncanonical autophagy through fluorescence microscopy.

Key words

Noncanonical autophagy LAP Phagocytosis ATG8 LC3 

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Copyright information

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

Authors and Affiliations

  • Elise Jacquin
    • 1
    • 2
  • Katherine Fletcher
    • 1
  • Oliver Florey
    • 1
    Email author
  1. 1.Signalling ProgrammeBabraham InstituteCambridgeUK
  2. 2.INSERM, U1231, Université de Bourgogne Franche ComtéDijonFrance

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