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Hedgehog Signaling Protocols

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Cover of 'Hedgehog Signaling Protocols'

Table of Contents

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    Book Overview
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    Chapter 1 Controlled Delivery of Sonic Hedgehog with a Heparin-Based Coacervate
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    Chapter 2 In Vivo Imaging of Hedgehog Transport in Drosophila Epithelia.
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    Chapter 3 Modeling Hedgehog Signaling Through Flux-Saturated Mechanisms
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    Chapter 4 Evaluating the Activity of Smoothened Toward G Proteins Using [ 35 S]Guanosine 5′-(3- O -thio)triphosphate ([ 35 S]GTPγS)
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    Chapter 5 Analysis of Smoothened Phosphorylation and Activation in Cultured Cells and Wing Discs of Drosophila
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    Chapter 6 Hedgehog Signaling Protocols
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    Chapter 7 Luciferase Reporter Assays to Study Transcriptional Activity of Hedgehog Signaling in Normal and Cancer Cells
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    Chapter 8 Measuring Expression Levels of Endogenous Gli Genes by Immunoblotting and Real-Time PCR
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    Chapter 9 Quantitative Immunoblotting of Endogenous Hedgehog Pathway Components
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    Chapter 10 Measuring Gli2 Phosphorylation by Selected Reaction Monitoring Mass Spectrometry
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    Chapter 11 Rapid Screening of Gli2/3 Mutants Using the Flp-In System
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    Chapter 12 Insights into Gli Factors Ubiquitylation Methods
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    Chapter 13 Determination of Acetylation of the Gli Transcription Factors
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    Chapter 14 Efficient Detection of Indian Hedgehog During Endochondral Ossification by Whole-Mount Immunofluorescence
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    Chapter 15 Methods for Detection of Ptc1-Driven LacZ Expression in Adult Mouse Skin
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    Chapter 16 Determination and Analysis of Cellular Metabolic Changes by Noncanonical Hedgehog Signaling.
Attention for Chapter 8: Measuring Expression Levels of Endogenous Gli Genes by Immunoblotting and Real-Time PCR
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Chapter title
Measuring Expression Levels of Endogenous Gli Genes by Immunoblotting and Real-Time PCR
Chapter number 8
Book title
Hedgehog Signaling Protocols
Published in
Methods in molecular biology, January 2015
DOI 10.1007/978-1-4939-2772-2_8
Pubmed ID
Book ISBNs
978-1-4939-2771-5, 978-1-4939-2772-2
Authors

Pawel Niewiadomski, Rajat Rohatgi

Abstract

Gli proteins are transcription factors that mediate the transcriptional effects of Hedgehog signaling in vertebrates. The activities of Gli2 and Gli3 are regulated primarily by posttranslational modifications, while Gli1 is mostly regulated at the transcriptional level. Detection of endogenous Gli proteins had been hampered by lack of good antibodies, but this problem has been mostly resolved in recent years. In this chapter we describe methods of detecting expression of endogenous Gli genes in whole-cell lysates and in subcellular fractions and also provide protocols for the measurement of Gli mRNA levels by quantitative real-time reverse transcriptase PCR (qPCR).

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Mendeley readers

The data shown below were compiled from readership statistics for 6 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 6 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 2 33%
Researcher 1 17%
Other 1 17%
Student > Master 1 17%
Unknown 1 17%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 3 50%
Agricultural and Biological Sciences 1 17%
Neuroscience 1 17%
Unknown 1 17%