Abstract
Necroptosis has been implicated as a critical cell death pathway in cancers, Alzheimer’s and other neurodegenerative diseases, and virus-infected cells. Necroptosis occurs when mixed-lineage kinase domain-like protein (MLKL) punctures the cytoplasmic membrane allowing a rapid influx of water leading to a loss of cellular integrity. As its role in human disease becomes apparent, methods identifying necroptosis will need to be further developed and optimized. Here we describe identification of necroptosis through quantifying cell death with pathway inhibitors and using western blots to identify end points of MLKL activation and protein-protein interactions leading to it.
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Acknowledgments
We would like to thank Nobuko Fukushima and James Bonner for technical assistance and Dr. Heather Koehler and Dr. Edward Mocarski for sharing expertise in necroptosis through consultation and protocol optimization.
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Cotsmire, S.M., Szczerba, M., Jacobs, B.L. (2021). Detecting Necroptosis in Virus-Infected Cells. In: Lucas, A.R. (eds) Viruses as Therapeutics. Methods in Molecular Biology, vol 2225. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1012-1_11
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DOI: https://doi.org/10.1007/978-1-0716-1012-1_11
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