The Hippo Pathway: Methods and Protocols

Front Matter ....Pages i-xiv
Front Matter ....Pages 1-1
 The Power of Drosophila Genetics: The Discovery of the Hippo Pathway  (Rewatee Gokhale, Cathie M. Pfleger)....Pages 3-26
 Drosophila Genetics: The Power of Genetic Mosaic Approaches  (Mardelle Atkins)....Pages 27-42
 Drosophila Genetics: Analysis of Tissue Growth in Adult Tissues  (Alexander D. Fulford, Paulo S. Ribeiro)....Pages 43-51
 Live Imaging of Hippo Pathway Components in Drosophila Imaginal Discs  (Jiajie Xu, Ting Su, Sherzod A. Tokamov, Richard G. Fehon)....Pages 53-59
 Localization of Hippo Signaling Components in Drosophila by Fluorescence and Immunofluorescence  (Cordelia Rauskolb, Kenneth D. Irvine)....Pages 61-73
 Bimolecular Fluorescence Complementation (BiFC) in Tissue Culture and in Developing Tissues of Drosophila to Study Protein-Protein Interactions  (Yurika Matsui, Zhi-Chun Lai)....Pages 75-85
Front Matter ....Pages 87-87
 Immunohistochemistry to Study YAP in Human Tissue Samples  (Franziska Haderk, Victor Olivas, Trever G. Bivona)....Pages 89-95
 Immunofluorescence Study of Endogenous YAP in Mammalian Cells  (Valentina Rausch, Carsten G. Hansen)....Pages 97-106
 Immunofluorescence Microscopy to Study Endogenous TAZ in Mammalian Cells  (Nathan M. Kingston, Andrew M. Tilston-Lunel, Julia Hicks-Berthet, Xaralabos Varelas)....Pages 107-113
 Nuclear/Cytoplasmic Fractionation to Study Hippo Effectors  (Maria Chatzifrangkeskou, Eric O’Neill)....Pages 115-119
 Luciferase Reporter Assays to Determine YAP/TAZ Activity in Mammalian Cells  (Sirio Dupont)....Pages 121-135
 Quantitative Real-Time PCR to Measure YAP/TAZ Activity in Human Cells  (Xiaolei Cao, Bin Zhao)....Pages 137-152
 HTRF® Total and Phospho-YAP (Ser127) Cellular Assays  (Diana Zindel, Claire Vol, Odile Lecha, Isabelle Bequignon, Merve Bilgic, Marion Vereecke et al.)....Pages 153-166
 Studying YAP-Mediated 3D Morphogenesis Using Fish Embryos and Human Spheroids  (Yoichi Asaoka, Hitoshi Morita, Hiroko Furumoto, Carl-Philipp Heisenberg, Makoto Furutani-Seiki)....Pages 167-181
 Regulation of YAP/TAZ Activity by Mechanical Cues: An Experimental Overview  (Sirio Dupont)....Pages 183-202
 CRISPR-Mediated Approaches to Regulate YAP/TAZ Levels  (Ryan J. Quinton, Neil J. Ganem)....Pages 203-214
 Hippo Pathway Regulation by Tyrosine Kinases  (Nina Reuven, Matan Shanzer, Yosef Shaul)....Pages 215-236
Front Matter ....Pages 237-237
 Structural and Biochemical Analyses of the Core Components of the Hippo Pathway  (Lisheng Ni, Xuelian Luo)....Pages 239-256
 Isothermal Titration Calorimetry Assays to Measure Binding Affinities In Vitro  (Kui Lin, Geng Wu)....Pages 257-272
 GST Pull-Down Assay to Measure Complex Formations  (Sun-Yong Kim, Toshio Hakoshima)....Pages 273-280
 Determining the Phosphorylation Status of Hippo Components YAP and TAZ Using Phos-tag  (Rui Chen, Steven W. Plouffe, Kun-Liang Guan)....Pages 281-287
 Quantifying the Kinase Activities of MST1/2  (Niamh A. O’Driscoll, David Matallanas)....Pages 289-304
 Measuring the Kinase Activities of the LATS/NDR Protein Kinases  (Alexander Hergovich)....Pages 305-317
 MST1/2 Kinase Assays Using Recombinant Proteins  (Marta Gomez, Yavuz Kulaberoglu, Alexander Hergovich)....Pages 319-331
Front Matter ....Pages 333-333
 Visualizing HIPPO Signaling Components in Mouse Early Embryonic Development  (Tristan Frum, Amy Ralston)....Pages 335-352
 The Hippo Signaling Pathway in Regenerative Medicine  (Lixin Hong, Yuxi Li, Qingxu Liu, Qinghua Chen, Lanfen Chen, Dawang Zhou)....Pages 353-370
Back Matter ....Pages 371-374