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. 2022:2459:169-177.
doi: 10.1007/978-1-0716-2144-8_16.

Imaging of Inflammasome Speck Formation in Living Cells

Lucas Secchim Ribeiro  1
Affiliations

Imaging of Inflammasome Speck Formation in Living Cells

Lucas Secchim Ribeiro. Methods Mol Biol. 2022.

Abstract

The detection of pathogen- or danger-associated molecular patterns during an inflammatory injury triggers the activation of cytosolic sensors known as inflammasomes. Once stimulated, these protein complexes can connect to the adaptor protein ASC, which in turn recruits the effector enzyme caspase-1, forming a polymeric structure known as ASC speck. This protein scaffold is responsible for processing cytokines of the IL-1 family into their active forms and evoking the cleavage of gasdermin D, ultimately leading to cell death by pyroptosis. Due to its micrometric size, the specks are used as a readout for inflammasome activation and for the better comprehension of this important immune pathway. In this chapter, a detailed protocol is presented for the study of the formation of inflammasome specks in living cells using confocal microscopy.

Keywords: ASC speck; Confocal microscopy; Inflammasome; Live-cell imaging; Pyroptosis.

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References

    1. Christgen S, Place DE, Kanneganti TD (2020) Toward targeting inflammasomes: insights into their regulation and activation. Cell Res 30(4):315–327. https://doi.org/10.1038/s41422-020-0295-8 - DOI - PubMed - PMC
    1. Swanson KV, Deng M, Ting JP (2019) The NLRP3 inflammasome: molecular activation and regulation to therapeutics. Nat Rev Immunol 19(8):477–489. https://doi.org/10.1038/s41577-019-0165-0 - DOI - PubMed - PMC
    1. Zheng D, Liwinski T, Elinav E (2020) Inflammasome activation and regulation: toward a better understanding of complex mechanisms. Cell Discov 6:36. https://doi.org/10.1038/s41421-020-0167-x - DOI - PubMed - PMC
    1. Broz P, Pelegrín P, Shao F (2020) The gasdermins, a protein family executing cell death and inflammation. Nat Rev Immunol 20(3):143–157. https://doi.org/10.1038/s41577-019-0228-2 - DOI - PubMed
    1. Hoss F, Rodriguez-Alcazar JF, Latz E (2017) Assembly and regulation of ASC specks. Cell Mol Life Sci 74(7):1211–1229. https://doi.org/10.1007/s00018-016-2396-6 - DOI - PubMed

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