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Detection of Apoptotic Versus Autophagic Cell Death by Flow Cytometry

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Book cover Programmed Cell Death

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1419))

Abstract

Different modes of regulated cell death (RCD) can be initiated by distinct molecular machineries and their morphological manifestations can be difficult to discriminate. Moreover, cells responding to stress often activate an adaptive response centered around autophagy, and whether such a response is cytoprotective or cytotoxic cannot be predicted based on morphological parameters only. Molecular definitions are therefore important to understand various RCD subroutines from a mechanistic perspective. In vitro, various forms of RCD including apoptosis and autophagic cell death can be easily discriminated from each other with assays that involve chemical or pharmacological interventions targeting key components of either pathway. Here, we detail a straightforward method to discriminate apoptosis from autophagic cell death by flow cytometry, based on the broad-spectrum caspase inhibitor Z-VAD-fmk and the genetic inhibition of ATG5.

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Acknowledgements

We are indebted to Dr. Ilio Vitale (Università di Roma “Tor Vergata”) for careful reading of the manuscript. Authors are supported by the Ligue contre le Cancer (équipe labellisée); Agence National de la Recherche (ANR); Association pour la recherche sur le cancer (ARC); Cancéropôle Ile-de-France; AXA Chair for Longevity Research; Institut National du Cancer (INCa); Fondation Bettencourt-Schueller; Fondation de France; Fondation pour la Recherche Médicale (FRM); the European Commission (ArtForce); the European Research Council (ERC); the LabEx Immuno-Oncology; the SIRIC Stratified Oncology Cell DNA Repair and Tumor Immune Elimination (SOCRATE); the SIRIC Cancer Research and Personalized Medicine (CARPEM); and the Paris Alliance of Cancer Research Institutes (PACRI).

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Authors and Affiliations

  1. Gustave Roussy Cancer Campus, Villejuif, France

    Valentina Sica, M. Chiara Maiuri & Lorenzo Galluzzi

  2. Equipe 11 labellisée par la Ligue Nationale contre le Cancer, Centre de Recherche des Cordeliers, 15, rue de l’Ecole de Médecine, 75006, Paris, France

    Valentina Sica, M. Chiara Maiuri, Guido Kroemer & Lorenzo Galluzzi

  3. INSERM, U1138, Paris, France

    Valentina Sica, M. Chiara Maiuri, Guido Kroemer & Lorenzo Galluzzi

  4. Faculté de Medicine, Université Paris Saclay/Paris XI, Le Kremlin-Bicêtre, France

    Valentina Sica

  5. Université Paris Descartes/Paris V, Sorbonne Paris Cité, Paris, France

    Valentina Sica, M. Chiara Maiuri, Guido Kroemer & Lorenzo Galluzzi

  6. Université Pierre et Marie Curie/Paris VI, Paris, France

    M. Chiara Maiuri, Guido Kroemer & Lorenzo Galluzzi

  7. Pôle de Biologie, Hôpital Européen Georges Pompidou AP-HP, Paris, France

    Guido Kroemer

  8. Metabolomics and Cell Biology Platforms, Gustave Roussy Cancer Campus, Villejuif, France

    Guido Kroemer

  9. Department of Women’s and Children’s Health, Karolinska University Hospital, Stockholm, Sweden

    Guido Kroemer

Authors
  1. Valentina Sica
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  2. M. Chiara Maiuri
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  3. Guido Kroemer
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  4. Lorenzo Galluzzi
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Corresponding authors

Correspondence to Guido Kroemer or Lorenzo Galluzzi .

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Editors and Affiliations

  1. Department of Biochemistry and Genetics, La Trobe Inst for Mol Sci, La Trobe Univ, Bundoora, Victoria, Australia

    Hamsa Puthalakath

  2. Department of Biochemistry and Genetics, La Trobe Inst for Mol Sci, La Trobe Univ, Bundoora, Victoria, Australia

    Christine J. Hawkins

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Sica, V., Maiuri, M.C., Kroemer, G., Galluzzi, L. (2016). Detection of Apoptotic Versus Autophagic Cell Death by Flow Cytometry. In: Puthalakath, H., Hawkins, C. (eds) Programmed Cell Death. Methods in Molecular Biology, vol 1419. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3581-9_1

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  • DOI: https://doi.org/10.1007/978-1-4939-3581-9_1

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