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Methods to Study the Effect of IKK Inhibition on TNF-Inducing Apoptosis and Necroptosis in Cultured Cells

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NF-κB Transcription Factors

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2366))

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Abstract

The engagement of TNF on TNFR can result in cell survival or cell death depending on the different complex formation downstream this interaction. Here we describe reagents and assay procedures that can be used to study caspase-independent cell death (necroptosis) in cultured cells, in response to pharmacological interventions with NF-kappaB and death inhibitors. We provide protocol to detect death-specific proteins using immunoblot and to dissect necrosome complex by sequential co-immunoprecipitation of death-specific components during necroptosis.

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Acknowledgments

We thank Dr. G. Courtois for MEF cells. The work was supported by the MIUR PRIN 2017WLKYAM_3 to A.P. and by France Incontinentia Pigmenti Foundation to M.V.U.

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Authors and Affiliations

  1. Institute of Genetics and Biophysics ‘Adriano Buzzati-Traverso’, Consiglio Nazionale delle Ricerche (CNR), Naples, Italy

    Alessandra Pescatore, Carmela Casale, Francesca Fusco & Matilde Valeria Ursini

Authors
  1. Alessandra Pescatore
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  2. Carmela Casale
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  3. Francesca Fusco
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  4. Matilde Valeria Ursini
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Corresponding author

Correspondence to Matilde Valeria Ursini .

Editor information

Editors and Affiliations

  1. Centre for Molecular Immunology and Inflammation, Department of Immunology and Inflammation, Imperial College London, London, UK

    Guido Franzoso

  2. Department of Biotechnological and Applied Clinical Sciences, University of L’Aquila, L’Aquila, Italy

    Francesca Zazzeroni

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Pescatore, A., Casale, C., Fusco, F., Ursini, M.V. (2021). Methods to Study the Effect of IKK Inhibition on TNF-Inducing Apoptosis and Necroptosis in Cultured Cells. In: Franzoso, G., Zazzeroni, F. (eds) NF-κB Transcription Factors. Methods in Molecular Biology, vol 2366. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1669-7_14

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  • DOI: https://doi.org/10.1007/978-1-0716-1669-7_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1668-0

  • Online ISBN: 978-1-0716-1669-7

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