This volume details up-to-date information and attempts to take the reader into the exciting realm of TGF- from the basic principles to the practical applications. Chapters provide basic introduction of TGF- signaling from the cell surface to the nucleus, methods and techniques for the investigation of TGF- signaling mechanism including receptors, Smads, intracellular kinases, microRNA, epigenetic regulation, post-translational regulations, non-Smad pathway; the physiological implications including those in cell cycle arrest, epithelial-mesenchymal transition, endothelial cells, adipogenesis, Th differentiation, stem cell, bone remodeling, ovary, zebrofish development, and frog animal capping; and the methodologies including metastasis imaging, 3D morphogenesis, membrane receptor quantification, conditional knockout, bone remodeling, kinase and phosphatase assays, BiFC interaction assays, and genome-wide siRNA screen. 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 key tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, TGF-Beta Signaling: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt

Regulation of TGF-b Receptors.- Determining TGF- Receptor Levels in the Cell Membrane.- Post-translational Modifications of TGF-b Receptors.- Production, Isolationand Structural Analysis of Ligands and Receptors of the TGF-b Superfamily.- Phosphorylation of Smads by Intracellular Kinases.- Analysis of Smad Phosphatase Activity in vitro.- Three-dimensional Mammary Epithelial Cell Morphogenesis Model for Analysis of TGF-b Signaling.- TGF- Signaling in Stem Cell Regulation.- Analysis of Epithelial-Mesenchymal Transition Induced by Transforming Growth Factor .- In vitro Th differentiation protocol.- Interrogating TGF-b Function and Regulation in Endothelial Cells.- Isolation and Manipulation of Adipogenic Cells to Assess TGF-b Superfamily Functions.- Imaging TGFb Signaling in Mouse Models of Cancer Metastasis.- Generation and Characterization of Smad7 Conditional Knockout Mice.- Monitoring Smad Activity in vivo Using the Xenopus Model System.- Animal Cap Assay for TGF-b Signaling.- Detection of Smad Signaling in Zebrafish Embryos.- Role of TGF- signaling in Coupling Bone Remodeling.- Studying the Functions of TGF- Signaling in the Ovary.- Quantitative Real-Time PCR Analysis of MicroRNAs and Their Precursors Regulated byTGF-b Signaling.- TGF- Regulated microRNAs and Their Function in Cancer Biology.- Epigenomic Regulation Of Smad1 Signaling During Cellular Senescence Induced by Ras Activation.- The Role of Ubiquitination to Determine Non-Smad Signaling Responses.- Genome-wide RNAi Screening to Dissect the TGF-b Signal Transduction Pathway.- Measuring TGF- Ligand Dynamics in Culture Medium.