UID:
edoccha_9958128828902883
Format:
1 online resource (379 p.)
ISBN:
0-12-801619-1
Series Statement:
Methods In Enzymology ; Volume 545
Content:
Regulated Cell Death Part A & Part B of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers research methods in apoptosis focusing on the important areas of intrinsic pathway, extrinsic pathway, caspases, cellular assays and post-apoptotic effects and model organisms; as well as topics on necroptosis and screening approaches.Continues the legacy of this premier serial with quality chapters authored by leaders in the field Covers research methods in biomineralization scienceRegulated Cell Death Part A & Par
Note:
Description based upon print version of record.
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Front Cover; Regulated Cell Death Part B: Necroptotic, Autophagic and other Non-apoptotic Mechanisms; Copyright; Contents; Contributors; Preface; Chapter One: Assays for Necroptosis and Activity of RIP Kinases; 1. Introduction; 1.1. Distinguishing features of necroptotic cell death; 1.2. Pathways and mediators of necroptosis; 2. Cellular Models of Necroptosis; 2.1. Cell types (Table 1.1); 2.2. Inducers of necroptosis; 2.3. Inhibitors of necroptosis; 3. Measurement of Necroptotic Cell Death
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3.1. Analysis of viability of FADD-deficient Jurkat cells treated with TNFα using CellTiter-Glo assay (Fig. 1.1)3.2. Determination of specific cell death using SYTOX Green assay (Fig. 1.2); 3.3. Annexin V/PI assay (Fig. 1.3); 3.4. Analysis of ROS increase (Fig. 1.4); 3.5. Mitochondrial membrane depolarization (Fig. 1.3); 3.6. Analysis of TNFα gene expression changes by qPCR (Fig. 1.5); 4. Recapitulation of RIP1 Kinase Expression in RIP1-Deficient Jurkat Cells; 4.1. Transient transfection (Fig. 1.6); 4.2. Generation of stable-inducible cell lines (Fig. 1.7)
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5. Analysis of Necrosome Complex Formation5.1. Immunoprecipitation of necrosome complex (Fig. 1.8); 5.2. Immunoprecipitation of TNFR1 complex; 5.3. Assessment of necrosome formation by fluorescence microscopy; 6. Endogenous RIPK Autophosphorylation Assays (Fig. 1.9); 7. Analysis of Recombinant RIPK1 Kinase Activity and Inhibition by Necrostatins; 7.1. Expression and purification of recombinant RIP1 and RIP3; 7.2. Kinase-Glo assay (Fig. 1.10); 7.3. HTRF KinEASE assay (Fig. 1.11); 7.4. Fluorescence polarization assay (Fig. 1.12); 7.5. Thermomelt assay (Fig. 1.13); 8. Conclusions
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AcknowledgmentsReferences; Chapter Two: IAP Family of Cell Death and Signaling Regulators; 1. Identification of IAPs, Structure, and Domain Function; 1.1. Discovery; 1.2. Domain structure-BIRs; 1.3. Domain structure-RING and UBA; 1.4. Domain structure-NACHT and enigmatic CARD; 2. IAP Proteins and Cell Death Pathways; 2.1. XIAP-Inhibitor of the intrinsic Bcl-2 blockable pathway; 2.2. XIAP-Caspase inhibitor; 2.3. Inhibition of cell death by c-IAP1 and c-IAP2; 2.4. IAP proteins and ubiquitin; 2.5. Regulation of signaling pathways by IAP proteins; 2.6. Targeting IAP proteins; References
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Chapter Three: Activation of the NLRP3 Inflammasome by Proteins That Signal for Necroptosis1. Introduction; 2. Altered Expression or Function of Enzymes That Control Induction of Necroptosis Results in Altered Generation of IL-1β...; 2.1. Generation of mouse bone marrow-derived DCs; 2.2. Use of transgenic mice to obtain DCs deficient in caspase-8 or RIPK3; 2.3. Knockdown of proteins signaling for necroptosis in DCs; 2.4. Induction of cytokines in DCs by agents inducing and activating the NLRP3 inflammasome; 2.5. Quantification of the induced cytokines
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3. Signaling Proteins Controlling Necroptosis Affect Assembly of the NLRP3 Inflammasome
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English
Additional Edition:
ISBN 0-12-801430-X
Language:
English
