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Methods for Assessing Autophagy and Autophagic Cell Death

  • Protocol
Autophagosome and Phagosome

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 445))

Summary

Autophagic (or type 2) cell death is characterized by the massive accumulation of autophagic vacuoles (autophagosomes) in the cytoplasm of cells that lack signs of apoptosis (type 1 cell death). Here we detail and critically assess a series of methods to promote and inhibit autophagy via pharmacological and genetic manipulations. We also review the techniques currently available to detect autophagy, including transmission electron microscopy, half-life assessments of long-lived proteins, detection of LC3 maturation/aggregation, fluorescence microscopy, and colocalization of mitochondrion- or endoplasmic reticulum–specific markers with lysosomal proteins. Massive autophagic vacuolization may cause cellular stress and represent a frustrated attempt of adaptation. In this case, cell death occurs with (or in spite of) autophagy. When cell death occurs through autophagy, on the contrary, the inhibition of the autophagic process should prevent cellular demise. Accordingly, we describe a strategy for discriminating cell death with autophagy from cell death through autophagy.

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Authors and Affiliations

  1. Unit “Apoptosis, Cancer and Immunity,”, INSERM, Villejuif, France

    Ezgi Tasdemir, Lorenzo Galluzzi, M. Chiara Maiuri, Alfredo Criollo, Ilio Vitale, Emilie Hangen, Nazanine Modjtahedi & Guido Kroemer

  2. Institut Gustave Roussy, Villejuif, France

    Ezgi Tasdemir, Lorenzo Galluzzi, M. Chiara Maiuri, Alfredo Criollo, Ilio Vitale, Emilie Hangen, Nazanine Modjtahedi & Guido Kroemer

  3. Faculté de Médecine–Université Paris-Sud, Villejuif, France

    Ezgi Tasdemir, Lorenzo Galluzzi, Alfredo Criollo, Ilio Vitale, Emilie Hangen, Nazanine Modjtahedi & Guido Kroemer

  4. Dipartimento di Farmacologia Sperimentale, Facoltà di Scienze Biotecnologiche–Università degli Studi di Napoli “Federico II”, Napoli, Italy

    M. Chiara Maiuri

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  1. Ezgi Tasdemir
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  2. Lorenzo Galluzzi
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  3. M. Chiara Maiuri
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  4. Alfredo Criollo
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  7. Nazanine Modjtahedi
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  8. Guido Kroemer
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Editors and Affiliations

  1. Department of Molecular Genetics and Microbiology, University of New Mexico Health Sciences Center, Albuquerque, NM, USA

    Vojo Deretic

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© 2008 Humana Press, a part of Springer Science+Business Media, LLC

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Tasdemir, E. et al. (2008). Methods for Assessing Autophagy and Autophagic Cell Death. In: Deretic, V. (eds) Autophagosome and Phagosome. Methods in Molecular Biology™, vol 445. Humana Press. https://doi.org/10.1007/978-1-59745-157-4_3

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  • DOI: https://doi.org/10.1007/978-1-59745-157-4_3

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-853-9

  • Online ISBN: 978-1-59745-157-4

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