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This volume explores techniques used to study and understand the latest research on pyroptosis. The chapters in this book cover topics such as experimental methods to induce pyroptosis via inflammasomes; approaches to identifying inflammasome activation in humans; ways to induce and detect pore formation; and techniques to evaluate cellular outcomes of pyroptotic cell death. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and thorough, Pyroptosis: Methods and Protocols is a valuable tool for researchers who want to expand their knowledge of this developing field. It will be especially beneficial for those working in the fields of immunology, cancer biology, microbiology, and cell biology.
List of contents
Molecular Mechanisms of Pyroptosis.- Assessment of NLRP3 Inflammasome Activation and NLRP3-NEK7 Complex Assembly.- Inducing Pyroptosis with FlaTox, RodTox, or NeedleTox.- Inducing Pyroptosis via the Pyrin Inflammasome.- Caspase-4 Activation and Recruitment to Intracellular Gram-Negative Bacteria.- CARD8 Inflammasome Activation by HIV-1 Protease.- Pyroptosis Markers in Human Primary Specimens: Quantification of Intracellular ASC Specks by Imaging Flow Cytometry and Extracellular Oxidized Mitochondrial by ELISA.- Identifying Inflammasome Activation in Human Tissues.- Analyzing Caspase-8-Dependent GSDMD Cleavage in Response to Yersinia Infection.- Biochemical Methods for Assessing Gasdermin D Inactivation in Macrophages.- Detection of Gasdermin C-Mediated Cancer Cell Pyroptosis.- Inducing Pyroptosis via Gasdermin B and Gasdermin E Cleavage.- Assessing Interleukin-1beta Activation during Pyroptosis.- Monitoring Calcium Fluxes and Lysosome Exocytosis during Pyroptosis.- Assessing Cytoskeletal Destruction During Pyroptosis.- Assessment of HMGB1 Release During Experimentally-Induced Pyroptosis.- Time-Lapse Imaging of Inflammasome-Dependent Cell Death and Extrusion in Enteroid-Derived Intestinal Epithelial Monolayers.
