Hedgehog Signaling Protocols

Controlled Delivery of Sonic Hedgehog with a Heparin-Based Coacervate.- In Vivo Imaging of Hedgehog Transport in Drosophila Epithelia.- Modeling Hedgehog Signaling Through Flux-Saturated Mechanism.- Evaluating the Activity of Smoothened Toward G Proteins Using [35S] Guanosine 5’-(3-O-thio)triphosphate ([35S]GTPγS).- Analysis of Smoothened Phosphorylation and Activation in Cultured Cells and Wing Discs of Drosophila.- Investigation of Protein-Protein Interactions and Conformational Changes in Hedgehog Signaling Pathway by FRET.- Luciferase Reporter Assays to Study Transcriptional Activity of HH Signaling in Normal and Cancer Cells.- Measuring Expression Levels of Endogenous Gli Genes by Immunoblotting and Real-Time PCR.- Quantitative Immunoblotting of Endogenous Hedgehog Pathway Components.- Measuring Gli2 Phosphorylation by Selected Reaction Monitoring Mass Spectrometry.- Rapid Screening of Gli2/3 Mutants Using the Flp-In System.- Insights into Gli Factors Ubiquitylation Methods.- Determination of Acetylation of the Gli Transcription Factors.- Efficient Detection of Indian Hedgehog During Endochondral Ossification by Whole Mount Immunofluorescence.- Methods for Detection of Ptc1-Driven LacZ Expression in Adult Mouse Skin.- Determination and Analysis of Cellular Metabolic Changes by Non-Canonical Hedgehog Signaling.