Hedgehog Signaling Protocols

Purifying the Hedgehog Protein and its Variants.- Application of Sonic Hedgehog to the Developing Chick Limb.- Manipulation of Hedgehog Signaling in Xenopus by Means of Embryo Microinjection and Application of Chemical Inhibitors.- Isolation of Rat Telencephalic Neural Explants to Assay Shh GABAergic Interneuron Differentiation-Inducing Activity.- Genetic Analysis of the Vertebrate Hedgehog-Signaling Pathway Using Muscle Cell Fate Specification in the Zebrafish Embryo.- Efficient Manipulation of Hedgehog/GLI Signaling Using Retroviral Expression Systems.- Cell Surface Marker and Cell Cycle Analysis, Hedgehog Signaling, and Flow Cytometry.- Detecting Tagged Hedgehog with Intracellular and Extracellular Immunocytochemistry for Functional Analysis.- Confocal Analysis of Hedgehog Morphogenetic Gradient Coupled with Fluorescent In Situ Hybridization of Hedgehog Target Genes.- RNAi in the Hedgehog Signaling Pathway: pFRiPE, a Vector for Temporally and Spatially Controlled RNAi in Drosophila.- Germline Clone Analysis for Maternally Acting Drosophila Hedgehog Components.- Clonal Analysis of Hedgehog Signaling in Drosophila Somatic Tissues.- GAL4/UAS Targeted Gene Expression for Studying Drosophila Hedgehog Signaling.- Biochemical Fractionation of Drosophila Cells.- Using Immunoprecipitation to Study Protein-Protein Interactions in the Hedgehog-Signaling Pathway.- Sequence Analyses to Study the Evolutionary History and Cis-Regulatory Elements of Hedgehog Genes.