Monitoring VEGF-Stimulated Calcium Ion Flux in Endothelial Cells.

Autor:	Critchley WR; School of Molecular & Cellular Biology, University of Leeds, Leeds, UK., Fearnley GWF; School of Molecular & Cellular Biology, University of Leeds, Leeds, UK., Abdul-Zani I; School of Molecular & Cellular Biology, University of Leeds, Leeds, UK., Molina-Paris C; School of Mathematics, University of Leeds, Leeds, UK., Bendtsen C; IMED Biotech Unit, AstraZeneca, Cambridge, UK., Zachary IC; Centre for Cardiovascular Biology and Medicine, University College London, London, UK., Harrison MA; School of Biomedical Sciences, University of Leeds, Leeds, UK., Ponnambalam S; School of Molecular & Cellular Biology, University of Leeds, Leeds, UK. s.ponnambalam@leeds.ac.uk.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2022; Vol. 2475, pp. 113-124.
DOI:	10.1007/978-1-0716-2217-9_7
Abstrakt:	The endothelial response to vascular endothelial growth factor A (VEGF-A) regulates many aspects of animal physiology in health and disease. Such VEGF-A-regulated phenomena include vasculogenesis, angiogenesis, tumor growth and progression. VEGF-A binding to receptor tyrosine kinases such as vascular endothelial growth factor receptor 2 (VEGFR2 ) activates multiple signal transduction pathways and changes in homeostasis, metabolism, gene expression, cell proliferation, migration, and survival. One such VEGF-A-regulated response is a rapid rise in cytosolic calcium ion levels which modulates different biochemical events and impacts on endothelial-specific responses. Here, we present a series of detailed and robust protocols for evaluating ligand-stimulated cytosolic calcium ion flux in endothelial cells. By monitoring an endogenous endothelial transcription factor (NFATc2 ) which displays calcium-sensitive redistribution, we can assess the relevance of cytosolic calcium to protein function. This protocol can be easily applied to both adherent and non-adherent cultured cells to evaluate calcium ion flux in response to exogenous stimuli such as VEGF-A. (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze:	MEDLINE
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