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This volume is divided in six section covering the most experimental approaches involved in the study of the unfolded protein response (UPR) pathway. Chapters detail determination of unfolded protein levels, methods to study UPR signal transmission, analysing the outcomes of the UPR pathway activation, UPR studies in mammalian models, UPR in alternative models, and UPR and disease. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.
Authoritative and cutting-edge, The Unfolded Protein Response: Methods and Protocols aims to describe key methods and approaches used in the study of the UPR pathway and its complex cellular implications.
Chapter 6 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.
List of contents
Measuring cysteine exposure in unfolded proteins with tetraphenylethene maleimide and its analogues.- Fluorescence-based biosensors for the detection of the unfolded protein response.- Indirect methods to measure unfolded proteins in living cells using fluorescent proteins.- A platform technology for monitoring the Unfolded Protein Response.- The use of fluorescent protein fusions to monitor the unfolded protein response and protein foldase-substrate interactions in plant protoplasts.- High-throughput analysis of protein turnover with tandem fluorescent protein timers.- Detection of HAC1 mRNA Splicing by RT-PCR in Saccharomyces cerevisiae.- Determination of protein translation during the activation of the UPR.- Determination of the stability and intracellular (intra-nuclear) targeting and recruitment of pre-HAC1 mRNA in the Saccharomyces cerevisiae during the activation of UPR.- Assays to study IRE1 activation and signaling.- Analysis of XBP1 contribution to hyperosmolarity-induced lipid synthesis.- Integrating an ER stress reporter for monitoring genome wide UPR-ER in budding yeast.- Analyze mouse knockout models of UPR pathway elements.- In vitro stimulation of IRE1a/XBP1-deficient B cells with LPS.- Detection of PERK signaling in the central nervous system.- Methods to study the mitochondrial unfolded protein response (UPRmt) in Caenorhabditis elegans.- Drosophila Unfolded Protein Response (UPR) assays in vitro and in vivo.- Protein preparation for proteomic analysis of the unfolded protein response in Arabidopsis thaliana.- Structure-based drug discovery of IRE1 modulators.- Chemical screening to detect ER stress regulators.
