Activation of the Non-canonical Inflammasome in Mouse and Human Cells

Jelena S Bezbradica, Rebecca C Coll, Dave Boucher*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


The non-canonical inflammasome is a signaling platform that allows for the detection of cytoplasmic lipopolysaccharides (LPS) in immune and non-immune cells. Upon detection of LPS, this inflammasome activates the signaling proteases caspase-4 and -5 (in humans) and caspase-11 (in mice). Inflammatory caspases activation leads to caspase self-processing and the cleavage of the pore-forming protein Gasdermin D (GSDMD). GSDMD N-terminal fragments oligomerize and form pores at the plasma membranes, leading to an inflammatory form of cell death called pyroptosis. Here, we describe a simple method to activate the non-canonical inflammasome in myeloid and epithelial cells and to measure its activity using cell death assay and immunoblotting.

Original languageEnglish
Pages (from-to)51-63
Number of pages13
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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