Platelet adhesion to podoplanin under flow is mediated by the receptor CLEC-2 and stabilised by Src/Syk-dependent platelet signalling

Leyre Navarro-Núñez; Alice Y. Pollitt; Kate Lowe; Arusa Latif; Gerard B. Nash; Steve P. Watson

doi:10.1160/TH14-09-0762

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2015; 113(05): 1109-1120
DOI: 10.1160/TH14-09-0762

Cellular Signalling and Proteolysis

Schattauer GmbH

Platelet adhesion to podoplanin under flow is mediated by the receptor CLEC-2 and stabilised by Src/Syk-dependent platelet signalling

Leyre Navarro-Núñez

¹Centre for Cardiovascular Sciences, Institute for Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK

,

Alice Y. Pollitt

¹Centre for Cardiovascular Sciences, Institute for Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK

,

Kate Lowe

¹Centre for Cardiovascular Sciences, Institute for Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK

,

Arusa Latif

¹Centre for Cardiovascular Sciences, Institute for Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK

,

Gerard B. Nash

¹Centre for Cardiovascular Sciences, Institute for Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK

,

Steve P. Watson

¹Centre for Cardiovascular Sciences, Institute for Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK

› Author Affiliations

Abstract

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Keywords

Platelet physiology - rheology - lymphatics - adhesion molecules - signal transduction

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Suppl. Video 1: Src and Syk inhibitors reduce stability of platelet adhesion to LECs. To analyse real-time adhesion of platelets to LECs, heparinized whole blood was incubated with FITC-conjugated anti-human CD41 and CD42b antibodies at 1/100 dilution and inhibitors or vehicle control as appropriate for 10 minutes and then perfused over LEC monolayers at 50 s^-1. Digital images were captured at 500 ms interval with 500 ms exposure time and 2×2 binning settings to create 1 minute videos. Upper left: vehicle; upper right: 9 µM integrilin; lower left: 10 µM dasatinib; lower right: 30 µM PRT060318.

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Suppl. Video 2: Src and Syk inhibitors reduce stability of platelet adhesion to hFcPDPN. To analyse real-time adhesion of platelets to hFcPDPN, heparinized whole blood was incubated with 4 µM DiOC6, 10µg/ml IV.3 antibody and inhibitors or vehicle control as appropriate for 10 minutes and then perfused over hFcPDPN-coated channels at 50 s^-1. Digital images were captured at 250 ms interval with 200 ms exposure time and 2×2 binning settings to create 1 minute videos. Upper left: vehicle; upper right: 9 µM integrilin; lower left: 10 µM dasatinib; lower right: 30 µM PRT060318.

References

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