Hedgehog Signaling Protocols

Cover......Page 1
Frontmatter......Page 2
1 Purifying the Hedgehog Protein and its Variants......Page 18
2 Application of Sonic Hedgehog to the Developing Chick Limb......Page 40
3 Manipulation of Hedgehog Signaling in Xenopus by Means of Embryo Microinjection and Application of Chemical Inhibitors......Page 51
4 Isolation of Rat Telencephalic Neural Explants to Assay Shh GABAergic Interneuron Differentiation-Inducing Activity......Page 62
5 Genetic Analysis of the Vertebrate Hedgehog-Signaling Pathway Using Muscle Cell Fate Specification in the Zebrafish Embryo......Page 69
6 Efficient Manipulation of Hedgehog/GLI Signaling Using Retroviral Expression Systems......Page 80
7 Cell Surface Marker and Cell Cycle Analysis, Hedgehog Signaling, and Flow Cytometry......Page 92
8 Detecting Tagged Hedgehog with Intracellular and Extracellular Immunocytochemistry for Functional Analysis......Page 104
9 Confocal Analysis of Hedgehog Morphogenetic Gradient Coupled with Fluorescent In Situ Hybridization of Hedgehog Target Genes......Page 117
10 RNAi in the Hedgehog Signaling Pathway: pFRiPE, a Vector for Temporally and Spatially Controlled RNAi in Drosophila......Page 126
11 Germline Clone Analysis for Maternally Acting Drosophila Hedgehog Components......Page 140
12 Clonal Analysis of Hedgehog Signaling in Drosophila Somatic Tissues......Page 156
13 GAL4/UAS Targeted Gene Expression for Studying Drosophila Hedgehog Signaling......Page 172
14 Biochemical Fractionation of Drosophila Cells......Page 213
15 Using Immunoprecipitation to Study Protein–Protein Interactions in the Hedgehog-Signaling Pathway......Page 224
16 Sequence Analyses to Study the Evolutionary History and Cis-Regulatory Elements of Hedgehog Genes......Page 239
Index......Page 259