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Analyzing Necroptosis Using an RIPK1 Kinase Inactive Mouse Model of TNF Shock

  • Matija Zelic
  • Michelle A. KelliherEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1857)

Abstract

The serine/threonine kinase RIPK1 has numerous biological and pathological functions, mediating prosurvival as well as prodeath apoptotic and necroptotic signaling pathways downstream of various receptors, including death receptors and Toll-like receptors (TLRs). RIPK1 has been implicated in various diseases, including ischemia–reperfusion injury and inflammatory bowel disease (IBD). The recent generation of RIPK1 kinase inactive mice has enabled us to genetically interrogate the role of RIPK1 kinase-mediated necroptosis in disease models. Here, we describe procedures utilizing kinase inactive Ripk1D138N/D138N mice to analyze necroptosis induction in vitro in bone-marrow derived macrophages (BMDMs) and in vivo in a murine model of TNF-induced shock.

Key words

Bone marrow-derived macrophages Lipopolysaccharide Caspase inhibitor TNF shock Cell death Necroptosis 

Notes

Acknowledgments

This work was supported by National Institutes of Health/National Institute of Allergy and Infectious Diseases Grant AI075118.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Molecular, Cell and Cancer BiologyUniversity of Massachusetts Medical SchoolWorcesterUSA

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