Influence of CD32 cross-linking on WASP and CrkL phosphorylation modulated by PI 3-kinase.

Anti-CD32 (3 μg/mL) was added to platelet suspensions for 10 minutes, followed by the addition of (Fab')2 goat antimouse IgG (30 μg/mL) for 5 minutes, as indicated. WASP (A) or CrkL (B) were purified from the soluble extracts by immunoprecipitation as described above, then immunoblotted with 4G10 (top panels) and anti-WASP or anti-CrkL (bottom panels). In panel B, the CrkL immunizing peptide (30 μg/mL) was added during precipitation as indicated. Arrows indicate bands of interest; HC, heavy chains of antibodies.