Fluorescence Correlation Spectroscopy Reveals Interaction of Some Microdomain-Associated Lipids with Cellular Focal Adhesion Sites
Abstract
:1. Introduction
2. Results
2.1. Localization of Focal Adhesion Sites of Cardiac Myofibroblasts
2.2. Distribution of Fluorescent Membrane Components in the Plasma Membrane of Cardiac Myofibroblasts
2.3. Diffusion of Fluorescent Lipids in the Plasma Membrane of Cardiac Myofibroblasts
2.4. Influence of Lipid Order on the Lipid Diffusion in Giant Unilamellar Vesicles (GUVs)
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. Identification of Myocytes and Fibroblasts by Immunostaining and Phase Contrast Microscopy
4.3. Transfection of Myofibroblasts with GFP-Vinculin
4.4. Lipids
4.5. Cell Staining Using Fusogenic Liposomes
4.6. Giant Unilamellar Vesicles (GUVs)
4.7. Supported Lipid Bilayer
4.8. Confocal Microscopy and Fluorescence Correlation Spectroscopy (FCS)
4.9. Statistical Analyses
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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D (μm2/s) | sdD (µm2/s) | Signi. D | NPart | sdPart | Signi. NPart | Nmeas | |||
---|---|---|---|---|---|---|---|---|---|
T-Test | Rank Sum Test | T-Test | Rank Sum Test | ||||||
BFL-SM (FA) | 1.0 | 0.3 | *** | *** | 23 | 17 | n.s. | n.s. | 54 |
BFL-SM (PM) | 1.3 | 0.3 | 21 | 17 | 54 | ||||
BFL-GM1 (FA) | 0.8 | 0.3 | *** | * | 38 | 30 | n.s. | n.s. | 37 |
BFL-GM1 (PM) | 1.0 | 0.4 | 41 | 54 | 37 | ||||
BFL-PC (FA) | 1.5 | 0.9 | n.s. | n.s. | 21 | 14 | n.s. | n.s. | 45 |
BFL-PC (PM) | 1.7 | 1.1 | 27 | 18 | 45 | ||||
TopChol (FA) | 3.7 | 1.4 | n.s. | n.s. | 29 | 15 | n.s. | n.s. | 35 |
TopChol (PM) | 3.7 | 1.6 | 32 | 18 | 35 | ||||
DiIC18(7) (FA) | 3.1 | 0.4 | n.s. | n.s. | 18 | 15 | n.s. | n.s. | 17 |
DiIC18(7) (PM) | 3.7 | 0.7 | 25 | 20 | 16 |
D (μm2/s) | sdD (µm2/s) | Signi. D | NPart | sdPart | Signi. NPart | Nmeas | |||
---|---|---|---|---|---|---|---|---|---|
T-test | Rank Sum Test | T-test | Rank Sum Test | ||||||
BFL-SM (Lo) | 0.9 | 0.2 | *** | *** | 72 | 59 | n.s. | n.s. | 10 |
BFL-SM (Ld) | 4.1 | 0.4 | 146 | 134 | 8 | ||||
BFL-GM1 (Lo) | 1.0 | 0.3 | *** | *** | 36 | 26 | n.s. | n.s. | 9 |
BFL-GM1 (Ld) | 4.2 | 0.4 | 48 | 23 | 10 | ||||
BFL-PC (Lo) | 1.1 | 0.3 | *** | *** | 12 | 7 | *** | *** | 12 |
BFL-PC (Ld) | 4.3 | 1.1 | 58 | 38 | 11 | ||||
TopChol (Lo) | 2.2 | 0.2 | *** | *** | 34 | 7 | *** | ** | 7 |
TopChol (Ld) | 10 | 0.9 | 62 | 11 | 7 | ||||
DiIC18(7) (Ld) | 6.8 | 0.4 | - | - | 3 | 1 | - | - | 7 |
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Kleusch, C.; Monzel, C.; Sridhar, K.C.; Hoffmann, B.; Csiszár, A.; Merkel, R. Fluorescence Correlation Spectroscopy Reveals Interaction of Some Microdomain-Associated Lipids with Cellular Focal Adhesion Sites. Int. J. Mol. Sci. 2020, 21, 8149. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21218149
Kleusch C, Monzel C, Sridhar KC, Hoffmann B, Csiszár A, Merkel R. Fluorescence Correlation Spectroscopy Reveals Interaction of Some Microdomain-Associated Lipids with Cellular Focal Adhesion Sites. International Journal of Molecular Sciences. 2020; 21(21):8149. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21218149
Chicago/Turabian StyleKleusch, Christian, Cornelia Monzel, Krishna Chander Sridhar, Bernd Hoffmann, Agnes Csiszár, and Rudolf Merkel. 2020. "Fluorescence Correlation Spectroscopy Reveals Interaction of Some Microdomain-Associated Lipids with Cellular Focal Adhesion Sites" International Journal of Molecular Sciences 21, no. 21: 8149. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21218149