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Quantification of Cell Cycle-Arresting Proteins

  • Oliver Kepp
  • Isabelle Martins
  • Laurie Menger
  • Mickaël Michaud
  • Sandy Adjemian
  • Abdul Qader Sukkurwala
  • Lorenzo Galluzzi
  • Guido KroemerEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 965)

Abstract

Cellular senescence, which can be defined as a stress response preventing the propagation of cells that have accumulated potentially oncogenic alterations, is invariably associated with a permanent cell cycle arrest. Such an irreversible blockage is mainly mediated by the persistent upregulation of one or more cyclin-dependent kinase inhibitors (CKIs), including (though not limited to) p16 INK4A and p21 CIP1 and p27 KIP1 . CKIs operate by binding to cyclin-dependent kinases (CDKs), de facto inhibiting their enzymatic activity. Here, we provide an immunoblotting-based method for the detection and quantification of CKIs in vitro and ex vivo, together with a set of guidelines for the interpretation of results.

Key words

ARF Cancer DNA damage INK p14ARF p57KIP2 

Notes

Acknowledgments

OK, LG and GK are especially grateful to Mr. Sundaramoorthy Balasubramanian, Senior Project Manager at SPi Global for assistance with the production of this book. GK is supported by the Ligue Nationale contre le Cancer (Equipe labellisée), Agence Nationale de la Recherche, Cancéropôle Ile-de-France, Fondation pour la Recherche Médicale, Institut National du Cancer, European Commission (Active p53, Apo-Sys, RIGHT, TransDeath, ChemoRes, ApopTrain), Fondation pour la Recherche Médicale, and the LabEx Immuno-Oncology.

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Copyright information

© Springer Science+Busincess Media, LLC 2013

Authors and Affiliations

  • Oliver Kepp
    • 1
    • 2
  • Isabelle Martins
    • 1
    • 2
  • Laurie Menger
    • 1
    • 2
  • Mickaël Michaud
    • 1
    • 2
  • Sandy Adjemian
    • 1
    • 2
  • Abdul Qader Sukkurwala
    • 1
    • 2
  • Lorenzo Galluzzi
    • 2
    • 3
  • Guido Kroemer
    • 1
    • 3
    • 4
    • 5
    • 6
    Email author
  1. 1.INSERM, U848VillejuifFrance
  2. 2.Institut Gustave RoussyVillejuifFrance
  3. 3.Université Paris DescartesParisFrance
  4. 4.Centre de Recherche des CordeliersParisFrance
  5. 5.Metabolomics PlatformInstitut Gustave RoussyVillejuifFrance
  6. 6.Pôle de BiologieHôpital Européen Georges Pompidou, AP-HPParisFrance

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