Related Experiment Video

Updated: Jan 25, 2026

Characterization of MLKL-mediated Plasma Membrane Rupture in Necroptosis
08:55

Characterization of MLKL-mediated Plasma Membrane Rupture in Necroptosis

Published on: August 7, 2018

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Necrosome Formation and Necroptosis in Experimental Cholestasis

Marta B Afonso1, Cecília M P Rodrigues2

  • 1Faculty of Pharmacy, Research Institute for Medicines (iMed.ULisboa), Universidade de Lisboa, Lisbon, Portugal.

Methods in Molecular Biology (Clifton, N.J.)
|April 25, 2019
PubMed

Insights

Necroptosis, a programmed cell death, is key in liver diseases like cholestasis. This study details methods to detect necroptosis in liver disease models, aiding research into its mechanisms.

Area of Science:

  • Hepatology
  • Cell Biology
  • Molecular Biology

Background:

  • Necroptosis is an increasingly recognized cell death pathway implicated in liver diseases, particularly cholestatic disorders.
  • This programmed cell death is activated by death receptor stimulation and relies on specific kinase activities and protein complex formation.
  • Understanding necroptosis is crucial for developing therapeutic strategies for liver pathologies.

Purpose of the Study:

  • To present a comprehensive set of methods for evaluating necroptosis.
  • To establish protocols for both in vitro and in vivo cholestasis models.
  • To facilitate the study of necroptosis in the context of liver disease.

Main Methods:

  • Detailed protocols for isolating total, soluble, and insoluble protein extracts from liver tissues and cell cultures.
  • In vitro kinase activity assays for receptor-interacting serine/threonine-protein kinase 3 (RIPK3).
  • Western blot analysis for detecting key necroptosis markers.

Main Results:

  • The study provides validated methods for assessing necroptosis.
  • The described techniques allow for detailed molecular analysis of necroptosis pathways.
  • The methods are applicable to various cholestatic liver disease models.

Conclusions:

  • The presented methodologies enable robust evaluation of necroptosis in liver disease research.
  • These methods will aid in elucidating the role of necroptosis in cholestasis.
  • This work provides essential tools for investigating targeted therapies for necroptosis-driven liver conditions.
Keywords:
Cell deathCell signalingCholestasisInsoluble protein fractionsMixed lineage kinase domain-like (MLKL)NecroptosisNecrosomeReceptor-interacting serine/threonine-protein kinase 3 (RIPK3)Tumor necrosis factor-α (TNF-α)Western blot

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