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Analysis of CDK Inhibitor Action on Mitochondria-Mediated Apoptosis

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Cyclin-Dependent Kinase (CDK) Inhibitors

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1336))

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Abstract

The role of cyclin-dependent kinase inhibitors (CDKIs) is to negatively regulate cyclin-dependent kinases as a mechanism of control of cell proliferation. As such, CDKIs are being used to induce apoptosis in cancer cells to prevent their excessive reproduction. This chapter describes procedures to study apoptosis induction upon treatment with any CDKI through the evaluation of morphological and functional mitochondrial alterations, in particular, how to measure the mitochondrial membrane potential (ΔΨm) using TMRE dye, determine the content of intracellular ATP, observe mitochondrial network morphology using HeLa cells stably expressing fluorescent reporter DsRed targeting mitochondrial matrix, observe ultrastructure of the organelle using transmission electron microscopy, and, finally, assure that mitochondrial outer membrane permeabilization takes place by assessing the subcellular localization of cyt C in HeLa cells stably expressing fluorescent cyt C-GFP.

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Authors and Affiliations

  1. Department of Anatomical Pathology, Pharmacology and Microbiology, University of Barcelona, Casanova 143, Barcelona, 08036, Spain

    Anna Gortat

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  1. Anna Gortat
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Correspondence to Anna Gortat .

Editor information

Editors and Affiliations

  1. Medicinal Chemistry, Centro de Investigación Príncipe Felipe, Valencia, Spain

    Mar Orzáez

  2. Medicinal Chemistry, Centro de Investigación Príncipe Felipe, Valencia, Spain

    Mónica Sancho Medina

  3. IBV-CSIC and Centro de Investigación Príncipe Felipe, Valencia, Spain

    Enrique Pérez-Payá

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Gortat, A. (2016). Analysis of CDK Inhibitor Action on Mitochondria-Mediated Apoptosis. In: Orzáez, M., Sancho Medina, M., Pérez-Payá, E. (eds) Cyclin-Dependent Kinase (CDK) Inhibitors. Methods in Molecular Biology, vol 1336. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2926-9_9

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  • DOI: https://doi.org/10.1007/978-1-4939-2926-9_9

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2925-2

  • Online ISBN: 978-1-4939-2926-9

  • eBook Packages: Springer Protocols

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