Critchley, WR orcid.org/0000-0002-2707-4080, Fearnley, GWF, Abdul-Zani, I et al. (5 more authors) (2022) Monitoring VEGF-Stimulated Calcium Ion Flux in Endothelial Cells. Methods in Molecular Biology, 2475. pp. 113-124. ISSN 1064-3745
Abstract
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.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2022. This is an author produced version of an article published in Methods in Molecular Biology. Uploaded in accordance with the publisher's self-archiving policy. |
Keywords: | Endothelial cells; Calcium; VEGF-A; VEGFR2; NFATc2; Human umbilical vein endothelial cells (HUVECs) |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Biomedical Sciences (Leeds) The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Cardiovascular Science (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 26 May 2022 13:17 |
Last Modified: | 21 Apr 2023 00:13 |
Status: | Published |
Publisher: | Springer |
Identification Number: | 10.1007/978-1-0716-2217-9_7 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:187354 |