Titlebook: NLR Proteins; Methods and Protocol Pablo Pelegrín,Francesco Di Virgilio Book 2023Latest edition The Editor(s) (if applicable) and The Autho

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Noncanonical NLRP3 Inflammasome Activation: Standard Protocols,, whose cleavage induces its pyroptotic activity. Thus, the analysis of cleaved GSDMD is an ideal pathway to study the noncanonical inflammasome. ELISA and immunoblot can be performed on cell culture supernatants or cell extracts.

Diatribe

Pyroptosis Induction and Visualization at the Single-Cell Level Using Optogenetics,e a protocol for the application of a novel optogenetics tool called NLS_PhoCl_N-GSDMD_mCherry that enables precise temporal and spatial pyroptosis induction in a confocal microscopy setup, followed by imaging of the cell death process and subsequent quantitative analysis of the experiment. This too

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Methods to Activate the NLRP3 Inflammasome, experimental approaches using primary murine bone marrow-derived macrophages (BMDMs) and bone marrow-derived dendritic cells (BMDCs), human peripheral blood mononuclear cells (PBMCs), as well as inflammasome-competent cell lines (HoxB8 and THP-1 cells). The protocol covers the use of a broad spectr