Main description:

This detailed book covers numerous expert methodologies to examine Hippo signaling on the structural, molecular, cellular, and organismal level. Beginning with a section on Drosophila genetics, the volume continues with parts exploring molecular and cell biological studies of the Hippo pathway, the use of structural biology and biochemistry, and a brief look at the Hippo pathway in mouse models. Written for the highly successful Methods in Molecular Biology series, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and practical, The Hippo Pathway: Methods and Protocols equips newcomers and specialists alike with key methodologies to accurately define the status of Hippo signaling in their experimental settings.

Contents:

Part I: The Hippo Pathway: Drosophila Genetics Lead the Way

1. The Power of Drosophila Genetics: The Discovery of the Hippo Pathway

Rewatee Gokhale and Cathie M. Pfleger

2. Drosophila Genetics: The Power of Genetic Mosaic Approaches

Mardelle Atkins

3. Drosophila Genetics: Analysis of Tissue Growth in Adult Tissues

Alexander D. Fulford and Paulo S. Ribeiro

4. Live Imaging of Hippo Pathway Components in Drosophila Imaginal Discs

Jiajie Xu, Ting Su, Sherzod A. Tokamov, and Richard G. Fehon

5. Localization of Hippo Signaling Components in Drosophila by Fluorescence and Immunofluorescence

Cordelia Rauskolb and Kenneth D. Irvine

6. Bimolecular Fluorescence Complementation (BiFC) in Tissue Culture and in Developing Tissues of Drosophila to Study Protein-Protein Interactions

Yurika Matsui and Zhi-Chun Lai

Part II: Molecular and Cell Biology Studies of the Hippo Pathway

7. Immunohistochemistry to Study YAP in Human Tissue Samples

Franziska Haderk, Victor Olivas, and Trever G. Bivona

8. Immunofluorescence Study of Endogenous YAP in Mammalian Cells

Valentina Rausch and Carsten G. Hansen

9. Immunofluorescence Microscopy to Study Endogenous TAZ in Mammalian Cells

Nathan M. Kingston, Andrew M. Tilston-Lunel, Julia Hicks-Berthet, and Xaralabos Varelas

10. Nuclear/Cytoplasmic Fractionation to Study Hippo Effectors

Maria Chatzifrangkeskou and Eric O'Neill

11. Luciferase Reporter Assays to Determine YAP/TAZ Activity in Mammalian Cells

Sirio Dupont

12. Quantitative Real-Time PCR to Measure YAP/TAZ Activity in Human Cells

Xiaolei Cao and Bin Zhao

13. HTRF (R) Total and Phospho-YAP(Ser127) Cellular Assays

Diana Zindel, Claire Vol, Odile Lecha, Isabelle Bequignon, Merve Bilgic, Marion Vereecke, Fabienne Charrier-Savournin, Maite Romier, Eric Trinquet, Jean-Philippe Pin, Julie Pannequin, Thomas Roux, Elodie Dupuis, and Laurent Prezeau

14. Studying YAP-Mediated 3D Morphogenesis Using Fish Embryos and Human Spheroids

Yoichi Asaoka, Hitoshi Morita, Hiroko Furumoto, Carl-Philipp Heisenberg, and Makoto Furutani-Seiki

15. Regulation of YAP/TAZ Activity by Mechanical Cues: An Experimental Overview

Sirio Dupont

16. CRISPR-Mediated Approaches to Regulate YAP/TAZ Levels

Ryan J. Quinton and Neil J. Ganem

17. Hippo Pathway Regulation by Tyrosine Kinases

Nina Reuven, Matan Shanzer, and Yosef Shaul

Part III: Structure Biology and Biochemistry to Study the Hippo Pathway

18. Structural and Biochemical Analyses of the Core Components of the Hippo Pathway

Lisheng Ni and Xuelian Luo

19. Isothermal Titration Calorimetry Assays to Measure Binding Affinities In Vitro

Kui Lin and Geng Wu

20. GST Pull-Down Assay to Measure Complex Formations

Sun-Yong Kim and Toshio Hakoshima

21. Determining the Phosphorylation Status of Hippo Components YAP and TAZ Using Phos-tag

Rui Chen, Steven W. Plouffe, and Kun-Liang Guan

22. Quantifying the Kinase Activities of MST1/2

Niamh A. O'Driscoll and David Matallanas

23. Measuring the Kinase Activities of the LATS/NDR Protein Kinases

Alexander Hergovich

24. MST1/2 Kinase Assays Using Recombinant Proteins

Marta Gomez, Yavuz Kulaberoglu, and Alexander Hergovich

Part IV: The Hippo Pathway and Mouse Models

25. Visualizing HIPPO Signaling Components in Mouse Early Embryonic Development

Tristan Frum and Amy Ralston

26. The Hippo Signaling Pathway in Regenerative Medicine

Lixin Hong, Yuxi Li, Qingxu Liu, Qinghua Chen, Lanfen Chen, and Dawang Zhou