Imaging of Intracellular and Plasma Membrane Pools of PI(4,5)P2 and PI4P in Human Platelets
Abstract
:1. Introduction
2. Materials and Methods
2.1. Antibodies
2.2. Human Platelet Isolation and Platelet Spreading
2.3. HEK293T and BALB3T3 Cells
2.4. Immunostaining and Confocal Fluorescence Microscopy
2.5. Statistical Analysis
3. Results
3.1. PI(4,5)P2 and PI4P Localize at Different Cellular Compartments in HEK293T and BALB3T3 Cells
3.2. Resting and Activated Platelets Readily Stain for the Intracellular Pools but Not PM Pools of PI(4,5)P2 and PI4P
3.3. PM Staining of PI(4,5)P2 and PI4P Can Be Improved by Decreasing the Permeabilization Time in Activated Platelets
3.4. Intracellular and PM PI(4,5)P2 and PI4P Can Be Modulated by OCRL and PI4KIIIα Inhibitors in Resting and Activated Platelets
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Bura, A.; Jurak Begonja, A. Imaging of Intracellular and Plasma Membrane Pools of PI(4,5)P2 and PI4P in Human Platelets. Life 2021, 11, 1331. https://doi.org/10.3390/life11121331
Bura A, Jurak Begonja A. Imaging of Intracellular and Plasma Membrane Pools of PI(4,5)P2 and PI4P in Human Platelets. Life. 2021; 11(12):1331. https://doi.org/10.3390/life11121331
Chicago/Turabian StyleBura, Ana, and Antonija Jurak Begonja. 2021. "Imaging of Intracellular and Plasma Membrane Pools of PI(4,5)P2 and PI4P in Human Platelets" Life 11, no. 12: 1331. https://doi.org/10.3390/life11121331