ELISA detection of wild-type and PAR-1 mutants on the surface of COS-7 cells
| Construct | Expression |
|---|---|
| % | |
| Wild-type | 100 |
| Deletions | |
| ΔNH3 (68–93) | 78 ± 5 |
| ΔEC3 (339–344) | 91 ± 17 |
| Point mutants | |
| I88A | 90 ± 14 |
| S89A | 93 ± 16 |
| L96A | 100 ± 6 |
| EC2DE (E241A, D256A, E260A, E264A) | 83 ± 10 |
| D256A | 101 ± 7 |
| D256N | NT |
| E260A | NT |
| E347A | 92 ± 12 |
| E347Q | 102 ± 10 |
| C175S | 57 ± 6 |
| C175S, C254S | 70 ± 9 |
COS-7 cells transiently expressing the wild-type or mutant receptors were incubated with thrombin for 10 min at 37°C. Expression levels were detected in fixed cells with a PAR-1 antibody as described. The expression levels are presented as a percentage of wild-type PAR-1 (100%). Data presented are mean of three independent experiments.
NT, not tested.