Part I. Wnt signaling pathway methods A: Canonical Wnt/FZD signaling Chapter 1. The Canonical Wnt/b -Catenin Signaling Pathway. Nick Barker Chapter 2. Isolation and application of bioactive Wnt proteins. Karl Willert Chapter 3: Purification and Wnt inhibitory activity of secreted Frizzled-related proteins. Vladimir Wolf, Yoshimi Endo and Jeffery S Rubin Chapter 4. Measuring GSK3 expression and activity in cells. Adam R Cole and Calum Sutherland Chapter 5.Inhibition of Glycogen Synthase Kinase-3. Andrei V Ougolkov and Daniel D Billadeau Chapter 6. Detection of cytoplasmic and nuclear localization of Adenomatous Polyposis Coli (APC) protein in cells. Mariana Brocardo and Beric R Henderson Chapter 7. Detection of b -catenin localization by immunohistochemistry. Nick Barker and Maaike van den Born Chapter 8. Assaying b -catenin/TCF transcription with b -catenin/TCF transcriptional-based reporter constructs. Travis L Biechele and Randall T Moon Chapter 9. Native promoter reporters validate transcriptional targets. Otto Schmalhofer, Simone Spaderna and Thomas Brabletz B: Non-canonical Signaling Chapter 10. b -Catenin-independent Wnt pathways: signals, core protein and effectors. Richard G James, William H Conrad and Randall T Moon Chapter 11. Image Analysis of Calcium Release Dynamics. Christina M Freisinger, Douglas W Houston and Diane Slusarski Chapter 12. Detecting PKC phosphorylation as part of the Wnt/calcium pathway in cutaneous melanoma. Samudra K Dissanayake and Ashani Weeraratna Chapter 13. Measuring CamK II activity in Xenopus embryos as a read-out for non-canonical Wnt signaling. Michael Kuhl and Petra Pandur Chapter 14. Analysis of Wnt7a-stimulated JNK activity and cJUN Phosphorylation in Non-Small Cell Lung Cancer Cells. Lynn E Heasley and Robert A Winn Chapter 15. ROCK enzymatic assay. John D Doran and Marc D Jacobs Chapter 16. Detection of Planar polarity proteins in mammalian cochlea. Mireille Montcouquiol and Nathalie Sans Part II. Mammalian Model systems for Wnt/FZD function Chapter 17. Proteomic analyses of protein complexes in the Wnt pathway. Stephane Angers Chapter 18. In situ hybridization to evaluate the expression of Wnt and Frizzled genes in mammalian tissues. Kestutis Planutis, Marina Planutiene and Randall F Holcombe Chapter 19. Assaying Wnt5a-mediated invasion in melanoma cells. Michael P. O'Connell, Amanda D. French, Poloko D. Leotlela and Ashani T. Weeraratna Chapter 20. Co-culture methodologies for the study of Wnt signals. Kestutis Planutis, Marina Planutiene and Randall F Holcombe Chapter 21. Analysis of Wnt/FZD-mediated signaling in a cell line model of colorectal cancer morphogenesis. Elizabeth Vincan, Robert H Whitehead and Maree C Faux Chapter 22. Analyzing tissue and gene function in intestinal organ culture. Helen E Abud. Heather M Young and Donald F Newgreen Chapter 23. The Genetics of Wnt Signaling During Early Mammalian Development. Terry Yamaguchi Chapter 24. Tissue-specific Transgenic, Conditional Knock-out and Knock-in Mice of Genes in the Canonical Wnt signaling Pathway. Koji Aoki and Makoto M Taketo
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.
Records our progress to date in understanding the intricacies of Wnt signalling
Includes assays for and models of both canonical and non-canonical Wnt signalling pathways
Contributing authors are either current or past members of the main laboratories that have shaped the Wnt field
Includes detailed methods as well as overviews, providing easy to follow protocols as well as a background/current status of the field