Figure 5. Botrocetin and shear cause GPIb–IX signalling and clearance of murine platelets.
(a) Illustration of the clearance study of botrocetin (Bc) injection. Purified botrocetin or PBS was injected intravenously into a C57BL/6J mouse, and blood was drawn at noted time points for platelet count. (b) Relative platelet count following botrocetin or PBS injection, with that before injection being 100%. Data are shown as mean±s.d. (n=6). (c) Illustration of the clearance study of in vitro sheared platelets. Murine platelets were collected from C57BL/6J mice, fluorescently labelled before being mixed with platelet-poor plasma (PPP) and botrocetin. The reconstituted PRP was treated with uniform shear of 18 dyn cm−2 before being either analysed by flow cytometry or being infused into a recipient mouse for clearance measurement. The PE-conjugated JON/A antibody selectively binds to the activated murine integrin αIIbβ3. (d) Botrocetin/shear treatment induces similar signalling in murine platelets. Quantificative plots of platelet signalling are plotted as described in Fig. 1b. (e) Clearance of in vitro sheared murine platelets, expressed as the percentage of CSFE-labelled platelets in total platelet population, after infusion. Data are shown as mean±s.d. (n=6). *P<0.05; **P<0.01; ***P<0.001.