This volume provides an updated collection of protocols for manipulating and studying VEGF signaling pathways in vitro and in vivo and aims to present a range of both firmly established and newly emerging technologies. Covering multiple model species, from mouse to zebrafish to human, the book explores the role of VEGF and VEGFR isoforms in exosomes, cultured cells, or in tissues, as well as robust cell assays for the investigation of basic angiogenic mechanisms and VEGF signaling in more complex cellular systems, amongst other subjects. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and up-to-date, VEGF Signaling: Methods and Protocols, Second Edition provides a useful tool for researchers in the vascular biology community and beyond in understanding the basic biology of VEGF signaling and in translating this research into the clinic.

Includes cutting-edge techniques Provides step-by-step detail essential for reproducible results Contains key implementation advice from the experts

Inhalt
Opposing Roles of Vascular Endothelial Growth Factor C in Metastatic Dissemination and Resistance to Radio/Chemotherapy: Discussion of Mechanisms and Therapeutic Strategies.- Identification of VEGF Isoforms in Mouse, Rat, and Zebrafish Using RT-qPCR.- Multi-Parameter Fluorescence-Activated Cell Sorting of Human Lymphatic Endothelial Cells.- Absolute Quantification of Plasma Membrane Receptors via Quantitative Flow Cytometry.- Measurement of VEGF Content in Exosomes and Subsequent Tumor Tubulogenesis and In Vivo Angiogenesis Functional Assays.- SH2-Domain Protein Isolation Using Synthetic Phosphorylated Peptides to Study VEGFR2 Signaling.- Monitoring VEGF-Stimulated Calcium Ion Flux in Endothelial Cells.- Co-Immunoprecipitation Assays.- Using Immortalized Endothelial Cells to Study the Roles of Adhesion Molecules in VEGF-Induced Signaling.- RNAscope for VEGF-A Detection in Human Tumor Bioptic Specimens.- Global MicroRNA Profiling of Vascular Endothelial Cells.- Endothelial Cell Tube Formation Assay: An In Vitro Model for Angiogenesis.- Use of a Thin Layer Assay for Assessing the Angiogenic Potential of Endothelial Cells In Vitro.- VEGF-A 165 -Induced Endothelial Cells Chemotactic Migration and Invasion Assays.- Measuring Mitochondrial Calcium Fluxes in Cardiomyocyte upon Mechanical Stretch-Induced Hypertrophy.- Simultaneous Measurement of Endothelial Cell Proliferation and Cell Cycle Stage Using Flow Cytometry.- Ex Vivo Mouse Aortic Ring Angiogenesis Assay.- Retinal Microvasculature-on-a-Chip for Modeling VEGF-Induced Permeability.- Preventing VEGF-Mediated Vascular Permeability by Experimentally Potentiating BBB Characteristics in Endothelial Cells.- The Embryonic Mouse Hindbrain and Postnatal Retina as In Vivo Models to Study Angiogenesis.- Evaluating VEGF-Induced Vascular Leakage Using the Miles Assay.- Modulation of VEGFA Signaling during Heart Regeneration in Zebrafish.- Three-Dimensional Visualization of Blood and Lymphatic Vessels in the Adult Zebrafish Heart by Chemical Clearing.- Fluorescence-Activated Cell Sorting and Quantitative Real-Time PCR to Reveal VEGF Expressing Macrophage Populations in the Zebrafish Larvae.- Assessing Molecular Regulation of Vascular Permeability Using a VEGF-Inducible Zebrafish Model.