Decrease in p85, GP Ib, and 14-3-3 association in the activated cytosol
| Immunoprecipitating antibody . | Immunoblotting antibody . | % Decrease in immunoreactive band after stimulation . |
|---|---|---|
| p85 | GP Ib | 35.26 ± 12.6 |
| (n = 4) | ||
| GP Ib | p85 | 45.32 ± 15.1 |
| (n = 8) | ||
| 14-3-3 | GP Ib | 37.02 ± 17.2 |
| (n = 7) | ||
| p85 | 50.37 ± 16.4 | |
| (n = 3) |
| Immunoprecipitating antibody . | Immunoblotting antibody . | % Decrease in immunoreactive band after stimulation . |
|---|---|---|
| p85 | GP Ib | 35.26 ± 12.6 |
| (n = 4) | ||
| GP Ib | p85 | 45.32 ± 15.1 |
| (n = 8) | ||
| 14-3-3 | GP Ib | 37.02 ± 17.2 |
| (n = 7) | ||
| p85 | 50.37 ± 16.4 | |
| (n = 3) |
Complex formation between the p85 subunit of PI 3-kinase, the GP Ib-IX receptor complex, and 14-3-3ζ was determined by immunoprecipitation with the indicated antibody from the Triton-soluble cytosolic fraction, followed by immunoblot analysis using antibodies to GP Ib or the p85 subunit. The percentage decrease in association between the immunoreactive species in thrombin-stimulated versus nonstimulated platelets was determined by densitometric analysis of the immunoreactive band. n, number of experiments.