VEGF Stimulates Activation of ERK5 in the Absence of C-Terminal Phosphorylation Preventing Nuclear Localization and Facilitating AKT Activation in Endothelial Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Agonists and Inhibitors
2.2. Antibodies and Reagents
2.3. Cell Culture
2.4. siRNA Knockdown
2.5. Agonist Stimulation
2.6. RIPA Protein Extraction and Western Blot
2.7. Immunofluorescence
2.8. qRT-PCR
2.9. MEF2 Reporter Assay
2.10. Statistical Analysis
3. Results
3.1. VEGF-Stimulated ERK5 Fails to Undergo C-Terminal Phosphorylation
3.2. VEGFR-2 Activation Suppresses ERK5 C-Terminal Phosphorylation in Endothelial Cells
3.3. VEGF-Stimulated ERK5 Regulates AKT Activity in HDMECs
3.4. VEGF-Stimulated ERK5 Does Not Undergo Nuclear Translocation and Co-Localises with Phosphorylated AKT
3.5. VEGF-Stimulated ERK5 Activity Does Not Regulate MEF2-Dependent Gene Expression in HDMECs
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Mondru, A.K.; Aljasir, M.A.; Alrumayh, A.; Nithianandarajah, G.N.; Ahmed, K.; Muller, J.; Goldring, C.E.P.; Wilm, B.; Cross, M.J. VEGF Stimulates Activation of ERK5 in the Absence of C-Terminal Phosphorylation Preventing Nuclear Localization and Facilitating AKT Activation in Endothelial Cells. Cells 2023, 12, 967. https://doi.org/10.3390/cells12060967
Mondru AK, Aljasir MA, Alrumayh A, Nithianandarajah GN, Ahmed K, Muller J, Goldring CEP, Wilm B, Cross MJ. VEGF Stimulates Activation of ERK5 in the Absence of C-Terminal Phosphorylation Preventing Nuclear Localization and Facilitating AKT Activation in Endothelial Cells. Cells. 2023; 12(6):967. https://doi.org/10.3390/cells12060967
Chicago/Turabian StyleMondru, Anil Kumar, Mohammad A. Aljasir, Ahmed Alrumayh, Gopika N. Nithianandarajah, Katie Ahmed, Jurgen Muller, Christopher E. P. Goldring, Bettina Wilm, and Michael J. Cross. 2023. "VEGF Stimulates Activation of ERK5 in the Absence of C-Terminal Phosphorylation Preventing Nuclear Localization and Facilitating AKT Activation in Endothelial Cells" Cells 12, no. 6: 967. https://doi.org/10.3390/cells12060967