Figure 1.
Loss of ITIM/ITSM phosphorylation prevents G6b-B–Shp1–Shp2 interaction. Flow cytometric (A) and western blot (B) analysis of G6b-B expression in platelets from WT and G6b-B diY/F mice. Mean ± SEM (n = 6) of G6b-FITC median fluorescence intensity (MFI), with rat IgG2A subtracted (Ai) and representative histogram (Aii). Representative blots (Bi) and quantification, normalized to tubulin reblots (mean ± SEM, n = 3) (Bii). (C) Lysates were prepared using washed platelets (5 × 108/mL) under basal conditions and 30 μg/mL collagen–stimulated conditions (90 seconds, 37°C, stirring at 1200 rpm) from WT and G6b-B diY/F mice. Coimmunoprecipitation of Shp1 and Shp2 was investigated by western blotting, following immunoprecipitation using anti–G6b-B or nonimmune rabbit polyclonal antibodies.