Gasdermin D Cleavage Assay Following Inflammasome Activation

Louisa Janice Kamajaya, Dave Boucher

Research output: Contribution to journalArticlepeer-review


Gasdermin D (GSDMD) is a recently identified pore-forming protein that is crucial for the execution of pyroptosis, a highly inflammatory form of cell death. GSDMD contains an N-terminal and a C-terminal domain that are separated by a proteolysis-sensitive linker. Upon cleavage of this linker by inflammasome-activated caspases, the N-terminal domain of GSDMD oligomerizes and forms pores at the plasma membrane, allowing cell swelling and subsequently membrane rupture to mediate pyroptosis. GSDMD is a key substrate of inflammatory caspases downstream of inflammasome activation and is driving various pathologies. Here, we describe a simple method to study GSDMD cleavage following canonical inflammasome activation in murine primary macrophages and neutrophils and human cell lines using immunoblotting.

Original languageEnglish
Pages (from-to)39-49
Number of pages11
JournalMethods in Molecular Biology
Publication statusPublished - 25 Feb 2022

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© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

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