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Wnt signaling. v. 1, Pathway methods and mammalian models / edited by Elizabeth Vincan. — Totowa, N.J. : Humana Press, c2008. – (58.17/M592/v.468) |
Contents
Contents
Preface
Contents
Contributors
PART I. WNT SIGNALING PATHWAY METHODS
SECTION A. CANONICAL WNT/FZD SIGNALING
1. The Canonical Wnt/β-Catenin Signalling Pathway
2. Isolation and Application of Bioactive Wnt Proteins
3. Purification and Wnt-Inhibitory Activities of Secreted Frizzled-Related Proteins
4. Measuring GSK3 Expression and Activity in Cells
5. Inhibition of Glycogen Synthase Kinase-3
6. Detection of Cytoplasmic and Nuclear Localization of Adenomatous Polyposis Coli (APC) Protein in Cells
7. Detection of β-Catenin Localization by Immunohistochemistry
8. Assaying β-Catenin/TCF Transcription with β-Catenin/TCF Transcription-Based Reporter Constructs
9. Native Promoter Reporters Validate Transcriptional Targets
SECTION B. NON-CANONICAL SIGNALING
10. β-Catenin-Independent Wnt Pathways: Signals, Core Proteins, and Effectors 131
11. Image Analysis of Calcium Release Dynamics 145
12. Detecting PKC Phosphorylation as Part of the Wnt/Calcium Pathway in Cutaneous Melanoma 157
13. Measuring CamKII Activity in Xenopus Embryos as a Read-out for Non-canonical Wnt Signaling 173
14. Analysis of Wnt 7a-Stimulated JNK Activity and cJun Phosphorylation in Non-Small Cell Lung Cancer Cells
15. ROCK Enzymatic Assay
16. Detection of Planar Polarity Proteins in Mammalian Cochlea 207
PART II. MAMMALIAN MODEL SYSTEMS FOR WNT/FZD FUNCTION
17. Proteomic Analyses of Protein Complexes in the Wnt Pathway
18. In Situ Hybridization to Evaluate the Expression of Wnt and Frizzled Genes in Mammalian Tissues
19. Assaying Wnt5A-Mediated Invasion in Melanoma Cells
20. Coculture Methodologies for the Study of Wnt Signals 255
21. Analysis of Wnt/FZD-Mediated Signalling in a Cell Line Model of Colorectal Cancer Morphogenesis 263
22. Analysing Tissue and Gene Function in Intestinal Organ Culture 275
23. Genetics of Wnt Signaling During Early Mammalian Development
24. Tissue-Specific Transgenic, Conditional Knockout and Knock-In Mice of Genes in the Canonical Wnt Signaling Pathway