WIP-WASp complex in platelets. (A) WT mouse platelets were activated with 3 μg/mL CRP or 0.5 U/mL thrombin for 0, 0.5, 1, and 2 minutes at 37°C as indicated. WASp was immunoprecipitated with mouse antibody B9. Immunoprecipitates were subjected to SDS-PAGE and probed with rabbit anti-WIP antibody C45 or anti-WASp antibody B4, as indicated. A control mouse IgG was used as negative control for specificity. (B) Platelet lysates before and after immunoprecipitation with anti-WASp antibody B9 or an isotype control were probed for WASp and WIP, as indicated. (C) Lysates corresponding to 4 × 106 WT, WASp KO, and WIP KO platelets were subjected to SDS-PAGE and probed with anti-WIP antibody C45 and anti-WASp antibody B4, as indicated. Anti-Syk probe was used as a control for loading. (D) WT and WIP KO platelets were probed with PE-labeled mouse anti-WASp antibody B9 (open) or an isotype control (shaded).
