PMID- 37074645
OWN - NLM
STAT- MEDLINE
DCOM- 20230421
LR  - 20230426
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 2641
DP  - 2023
TI  - Analyzing Caspase-8-Dependent GSDMD Cleavage in Response to Yersinia Infection.
PG  - 115-124
LID - 10.1007/978-1-0716-3040-2_9 [doi]
AB  - 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.
CI  - (c) 2023. The Author(s), under exclusive license to Springer Science+Business 
      Media, LLC, part of Springer Nature.
FAU - Chan, Felicia Hui Min
AU  - Chan FHM
AD  - Immunology Translational Research Programme, Department of Microbiology and 
      Immunology, Yong Loo Lin School of Medicine, National University of Singapore, 
      Singapore, Singapore.
AD  - Immunology Programme, Life Sciences Institute, National University of Singapore, 
      Singapore, Singapore.
FAU - Chen, Kaiwen W
AU  - Chen KW
AD  - Immunology Translational Research Programme, Department of Microbiology and 
      Immunology, Yong Loo Lin School of Medicine, National University of Singapore, 
      Singapore, Singapore. Kaiwen.chen@nus.edu.sg.
AD  - Immunology Programme, Life Sciences Institute, National University of Singapore, 
      Singapore, Singapore. Kaiwen.chen@nus.edu.sg.
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - EC 3.4.22.- (Caspase 8)
RN  - 0 (Apoptosis Regulatory Proteins)
RN  - EC 3.4.22.36 (Caspase 1)
RN  - 0 (Inflammasomes)
SB  - IM
MH  - Mice
MH  - Animals
MH  - Caspase 8/metabolism
MH  - *Apoptosis/physiology
MH  - *Yersinia Infections/metabolism
MH  - Apoptosis Regulatory Proteins/metabolism
MH  - Macrophages/metabolism
MH  - Caspase 1/metabolism
MH  - Inflammasomes/metabolism
OTO - NOTNLM
OT  - Caspase-8
OT  - Gasdermin D
OT  - Macrophage
OT  - Pyroptosis
OT  - Yersinia
EDAT- 2023/04/19 12:42
MHDA- 2023/04/21 06:41
CRDT- 2023/04/19 11:24
PHST- 2023/04/21 06:41 [medline]
PHST- 2023/04/19 12:42 [pubmed]
PHST- 2023/04/19 11:24 [entrez]
AID - 10.1007/978-1-0716-3040-2_9 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2023;2641:115-124. doi: 10.1007/978-1-0716-3040-2_9.