Analyzing Caspase-8-Dependent GSDMD Cleavage in Response to Yersinia Infection.
Caspase-8
Gasdermin D
Macrophage
Pyroptosis
Yersinia
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2023
2023
Historique:
medline:
21
4
2023
pubmed:
19
4
2023
entrez:
19
04
2023
Statut:
ppublish
Résumé
Caspase-8 is best known to drive an immunologically silent form of cell death known as apoptosis. However, emerging studies revealed that upon pathogen inhibition of innate immune signalling, such as during Yersinia infection in myeloid cells, caspase-8 associates with RIPK1 and FADD to trigger a proinflammatory death-inducing complex. Under such conditions, caspase-8 cleaves the pore-forming protein gasdermin D (GSDMD) to trigger a lytic form of cell death, known as pyroptosis. Here, we describe our protocol to activate caspase-8-dependent GSDMD cleavage following Yersinia pseudotuberculosis infection in murine bone marrow-derived macrophages (BMDMs). Specifically, we describe protocols on harvesting and plating of BMDM, preparation of type 3 secretion system-inducing Yersinia, macrophage infection, lactate dehydrogenase (LDH) release assay, and Western blot analysis.
Identifiants
pubmed: 37074645
doi: 10.1007/978-1-0716-3040-2_9
doi:
Substances chimiques
Caspase 8
EC 3.4.22.-
Apoptosis Regulatory Proteins
0
Caspase 1
EC 3.4.22.36
Inflammasomes
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
115-124Informations de copyright
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
Références
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