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Hedgehog Signaling Protocols

  • Book
  • © 2015

Overview

Editors:
  1. Natalia A. Riobo

    1. Department of Biochemistry and Molecular Biology, Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, USA

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

  • Includes cutting-edge methods and protocols
  • Provides step-by-step detail essential for reproducible results
  • Contains key notes and implementation advice from the experts
  • Includes supplementary material: sn.pub/extras

Part of the book series: Methods in Molecular Biology (MIMB, volume 1322)

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Table of contents (16 protocols)

  1. Front Matter

    Pages i-xvi
    Download chapter PDF

  2. Controlled Delivery of Sonic Hedgehog with a Heparin-Based Coacervate

    • Noah Ray Johnson, Yadong Wang
    Pages 1-7

  3. In Vivo Imaging of Hedgehog Transport in Drosophila Epithelia

    • Irene Seijo-Barandiarán, Isabel Guerrero, Marcus Bischoff
    Pages 9-18

  4. Modeling Hedgehog Signaling Through Flux-Saturated Mechanisms

    • Óscar Sánchez, Juan Calvo, Carmen Ibáñez, Isabel Guerrero, Juan Soler
    Pages 19-33

  5. Evaluating the Activity of Smoothened Toward G Proteins Using [35S]Guanosine 5′-(3-O-thio)triphosphate ([35S]GTPγS)

    • David R. Manning, Feng Shen, Natalia A. Riobo
    Pages 35-44

  6. Analysis of Smoothened Phosphorylation and Activation in Cultured Cells and Wing Discs of Drosophila

    • Kai Jiang, Jianhang Jia
    Pages 45-60

  7. Investigation of Protein–Protein Interactions and Conformational Changes in Hedgehog Signaling Pathway by FRET

    • Lin Fu, Xiangdong Lv, Yue Xiong, Yun Zhao
    Pages 61-70

  8. Luciferase Reporter Assays to Study Transcriptional Activity of Hedgehog Signaling in Normal and Cancer Cells

    • Silvia Pandolfi, Barbara Stecca
    Pages 71-79

  9. Measuring Expression Levels of Endogenous Gli Genes by Immunoblotting and Real-Time PCR

    • Pawel Niewiadomski, Rajat Rohatgi
    Pages 81-92

  10. Quantitative Immunoblotting of Endogenous Hedgehog Pathway Components

    • Shohreh F. Farzan, David J. Robbins
    Pages 93-104

  11. Measuring Gli2 Phosphorylation by Selected Reaction Monitoring Mass Spectrometry

    • Robert Ahrends, Pawel Niewiadomski, Mary N. Teruel, Rajat Rohatgi
    Pages 105-123

  12. Rapid Screening of Gli2/3 Mutants Using the Flp-In System

    • Pawel Niewiadomski, Rajat Rohatgi
    Pages 125-130

  13. Insights into Gli Factors Ubiquitylation Methods

    • Paola Infante, Romina Alfonsi, Lucia Di Marcotullio
    Pages 131-146

  14. Determination of Acetylation of the Gli Transcription Factors

    • Sonia Coni, Laura Di Magno, Gianluca Canettieri
    Pages 147-156

  15. Efficient Detection of Indian Hedgehog During Endochondral Ossification by Whole-Mount Immunofluorescence

    • João Francisco Botelho, Daniel Smith-Paredes, Verónica Palma A.
    Pages 157-166

  16. Methods for Detection of Ptc1-Driven LacZ Expression in Adult Mouse Skin

    • Donna M. Brennan-Crispi, Mỹ G. Mahoney, Natalia A. Riobo
    Pages 167-185

  17. Determination and Analysis of Cellular Metabolic Changes by Noncanonical Hedgehog Signaling

    • Raffaele Teperino, John Andrew Pospisilik
    Pages 187-198

  18. Back Matter

    Pages 199-200
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Keywords

About this book

The second edition of this volume provides novel protocols for the study of newly discovered function and modifications of components of the Hh pathways. The first few chapters discuss methods for stable delivery of soluble Hh proteins in cell cultures that can be adapted for in vivo studies, and mathematical models for studying Hh proteins gradient formation in tissues. The next few chapters explore studies in cell culture using fibroblasts that respond readily to Hh proteins. The book goes on to present protocols for evaluation of transcriptional activity of the Gli transcription factors in cell cultures and determine the expression level of endogenous Gli proteins. The final chapters talk about metabolic effects of noncanonical Hh signaling. Written in the highly successful Methods in Molecular Biology series format, chapters include 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 authoritative, Hedgehog Signaling Protocols, Second Edition presents researchers with a comprehensive understanding of the Hh pathway and its function in development and diseases.

Editors and Affiliations

  • Department of Biochemistry and Molecular Biology, Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, USA

    Natalia A. Riobo

Bibliographic Information

  • Book Title: Hedgehog Signaling Protocols

  • Editors: Natalia A. Riobo

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-4939-2772-2

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media New York 2015

  • Hardcover ISBN: 978-1-4939-2771-5Published: 21 July 2015

  • Softcover ISBN: 978-1-4939-4922-9Published: 15 October 2016

  • eBook ISBN: 978-1-4939-2772-2Published: 15 July 2015

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 2

  • Number of Pages: XVI, 200

  • Number of Illustrations: 14 b/w illustrations, 28 illustrations in colour

  • Topics: Protein Science

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