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E-raamat: Inflammasome: Methods and Protocols

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 Inflammasomes have a seemingly simple architecture. A sensor molecule of the NLR or PYHIN protein family recruits the common adapter molecule apoptosis-associated speck-like protein containing a CARD (ASC). Even so, inflammasome activation is highly regulated at multiple steps and precise details of how inflammasomes are activated at the molecular level remain largely unknown. In The Inflammasome: Methods and Protocols, expert researchers in the field detail methods and protocols to study multiple aspects of inflammasome function. These methods include the generation of inflammasome stimuli, perturbance of cellular homeostasis, assessment of ASC speck formation, monitoring of caspase-1 activity and processing, activation of IL-1 cytokines, and analysis of inflammasome assembly. 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 key tips on troubleshooting and avoiding known pitfalls.
Preface v
Contributors ix
1 Assessing β-Amyloid-Induced NLRP3 Inflammasome Activation in Primary Microglia
1(8)
Mareike Schnaars
Hannes Beckert
Annett Halle
2 Activating the NLRP3 Inflammasome Using the Amyloidogenic Peptide IAPP
9(10)
Clara Westwell-Roper
Aisling Dunne
Man Lyang Kim
C. Bruce Verchere
Seth L. Masters
3 Assessment and Quantification of Crystal-Induced Lysosomal Damage
19(10)
Peter Duewell
Eicke Latz
4 Assessment of Inflammasome Activation in Primary Human Immune Cells
29(12)
Theo S. Plantinga
Leo A.B. Joosten
Mihai G. Netea
5 NLRP3 Inflammasome Activation and Cytotoxicity Induced by Particulate Adjuvants
41(24)
Marie Yang
Claire H.A. Hearnden
Ewa Oleszycka
Ed C. Lavelle
6 Measuring Inflammasome Activation in Response to Bacterial Infection
65(20)
Petr Broz
Denise M. Monack
7 Detection of Pyroptosis by Measuring Released Lactate Dehydrogenase Activity
85(6)
Manira Rayamajhi
Yue Zhang
Edward A. Miao
8 ASC Speck Formation as a Readout for Inflammasome Activation
91(12)
Andrea Stutz
Gabor L. Horvath
Brian G. Monks
Eicke Latz
9 Immunoblotting for Active Caspase-1
103(14)
Christopher Jakobs
Eva Bartok
Andrej Kubarenko
Franz Bauernfeind
Veit Hornung
10 Inflammasome Activation and Inhibition in Primary Murine Bone Marrow-Derived Cells, and Assays for IL-1,α IL-1β, and Caspase-1
117(20)
Katharina S. Schneider
Christina J. Thomas
Olaf Groß
11 Reconstituting the NLRP1 Inflammasome In Vitro
137(16)
Benjamin Faustin
John C. Reed
12 Assessing ATP Binding and Hydrolysis by NLR Proteins
153(16)
Jinyao Mo
Joseph A. Duncan
13 Blue Native Polyacrylamide Gel Electrophoresis to Monitor Inflammasome Assembly and Composition
169(16)
Eric M. Kofoed
Russell E. Vance
14 Analysis of Microbiota Alterations in Inflammasome-Deficient Mice
185(10)
Eran Elinav
Christoph A. Thaiss
Richard A. Flavell
15 Quantification of Adipose Tissue Leukocytosis in Obesity
195(16)
Ryan Grant
Yun-Hee Youm
Anthony Ravussin
Vishwa Deep Dixit
16 In Vivo Evaluation of Neutrophil Recruitment in Response to Sterile Particulates
211(12)
Tamiko Yanagida
Kaoru Orihashi
Hajime Kono
Index 223