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. Author manuscript; available in PMC: 2015 Aug 1.
Published in final edited form as: Vascul Pharmacol. 2014 Jan 25;62(2):63–71. doi: 10.1016/j.vph.2014.01.005

Figure 4. Heparin and ODSH protect PAR-1 from cleavage by thrombin.

Figure 4

Panels A, B. HPAEC were pre-treated with heparin (A) or ODSH (B) (50 µg/ml, 40 min), washed with fresh serum-free cell culture medium to remove unbound heparins, then challenged with thrombin (8 nM, 10 min) and analyzed by Western blotting using antibody specifically recognizing thrombin-cleaved PAR-1. Statistical analyses of the experimental data expressed in relative units are presented as a mean of three individual experiments ± SE. *P < 0.01 versus cells treated with thrombin alone. Panel C. Heparin and ODSH affect cell surface expression of PAR-1. Exposure to thrombin, heparin or ODSH alters cell surface expression of PAR-1 as revealed by immunofluorescence. Cells were incubated with thrombin (20 ng/ml) or heparin (50 µg/ml) or ODSH (50 µg/ml) for 30 min prior to immunostaining with anti-PAR-1 antibody. Images of control, thrombin-, heparin- and ODSH-treated EC are shown. Results are representative of a minimum of 3 individual experiments.