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Since their discovery, Wnt signaling molecules have been shown to control key events in embryogenesis, to maintain tissue homeostasis in the adult and, when aberrantly activated, to promote human degenerative diseases and cancer, thus making them a vital area of study. Wnt Signaling: Methods and Protocols examines both biochemical assays and vertebrate and invertebrate model systems to provide a point of reference to current molecular protocols and the diverse model systems employed to study this important signaling pathway. In Volume 1, Pathway Methods and Mammalian Models, assays to measure activation of the diverse Wnt signaling pathways are explored, along with a selection of models and strategies used to study mammalian Wnt/FZD fuction, from protein-protein interaction and simple cell line models to organoid cultures and mouse models. As a volume in the highly successful Methods in Molecular Biology(TM) series, chapters contain readily reproducible laboratory protocols, complete with lists of necessary equipment and reagents and the Notes section, which reveals helpful troubleshooting tips.
Comprehensive and cutting-edge, Wnt Signaling: Methods and Protocols collects the expertise and knowledge of many leaders in the field to produce this invaluable two-volume resource.
List of contents
Wnt Signaling Pathway Methods.- The Canonical Wnt/?-Catenin Signalling Pathway.- Isolation and Application of Bioactive Wnt Proteins.- Purification and Wnt-Inhibitory Activities of Secreted Frizzled-Related Proteins.- Measuring GSK3 Expression and Activity in Cells.- Inhibition of Glycogen Synthase Kinase-3.- Detection of Cytoplasmic and Nuclear Localization of Adenomatous Polyposis Coli (APC) Protein in Cells.- Detection of ?-Catenin Localization by Immunohistochemistry.- Assaying ?-Catenin/TCF Transcription with ?-Catenin/TCF Transcription-Based Reporter Constructs.- Native Promoter Reporters Validate Transcriptional Targets.- ?-Catenin-Independent Wnt Pathways: Signals, Core Proteins, and Effectors.- Image Analysis of Calcium Release Dynamics.- Detecting PKC Phosphorylation as Part of the Wnt/Calcium Pathway in Cutaneous Melanoma.- Measuring CamKII Activity in Xenopus Embryos as a Read-out for Non-canonical Wnt Signaling.- Analysis of Wnt7a-Stimulated JNK Activity and cJun Phosphorylation in Non-Small Cell Lung Cancer Cells.- ROCK Enzymatic Assay.- Detection of Planar Polarity Proteins in Mammalian Cochlea.- Mammalian Model Systems for Wnt/FZD Function.- Proteomic Analyses of Protein Complexes in the Wnt Pathway.- In Situ Hybridization to Evaluate the Expression of Wnt and Frizzled Genes in Mammalian Tissues.- Assaying Wnt5A-Mediated Invasion in Melanoma Cells.- Coculture Methodologies for the Study of Wnt Signals.- Analysis of Wnt/FZD-Mediated Signalling in a Cell Line Model of Colorectal Cancer Morphogenesis.- Analysing Tissue and Gene Function in Intestinal Organ Culture.- Genetics of Wnt Signaling During Early Mammalian Development.- Tissue-Specific Transgenic, Conditional Knockout and Knock-In Mice of Genes in the Canonical Wnt Signaling Pathway.
Summary
Since their discovery, Wnt signaling molecules have been shown to control key events in embryogenesis, to maintain tissue homeostasis in the adult and, when aberrantly activated, to promote human degenerative diseases and cancer, thus making them a vital area of study. Wnt Signaling: Methods and Protocols examines both biochemical assays and vertebrate and invertebrate model systems to provide a point of reference to current molecular protocols and the diverse model systems employed to study this important signaling pathway. In Volume 1, Pathway Methods and Mammalian Models, assays to measure activation of the diverse Wnt signaling pathways are explored, along with a selection of models and strategies used to study mammalian Wnt/FZD fuction, from protein-protein interaction and simple cell line models to organoid cultures and mouse models. As a volume in the highly successful Methods in Molecular Biology™ series, chapters contain readily reproducible laboratory protocols, complete with lists of necessary equipment and reagents and the Notes section, which reveals helpful troubleshooting tips.
Comprehensive and cutting-edge, Wnt Signaling: Methods and Protocols collects the expertise and knowledge of many leaders in the field to produce this invaluable two-volume resource.
