Fig. 3

The defect of spreading of Tyro3^−/− and Axl^−/− platelets stimulated with poly(PHG) on a fibrinogen-coated surface. Washed platelets from wild-type, Tyro3^−/−, Axl^−/−, or Mertk^−/− mice were seeded on coverslips coated with 10 μg mL^− 1 fibrinogen in the presence or absence of 5 ng mL^− 1 poly(PHG) for 30 min. After washing, adherent platelets were fixed in 4% paraformaldehyde at room temperature for 30 min and stained with TRITC-conjugated phalloidin (1:500) containing 0.1% Triton X-100 for 2 h. The samples were observed under a fluorescent microscope and photographed using the Olympus FSX100. Representative images are shown (a i) and histograms represent mean ± SEM of five independent experiments (a ii). Expression levels of glycoproteins (αIIbβ3, GPIb, or GPVI) on platelets from wild-type, Tyro3^−/−, Axl^−/−, and Mertk^−/− mice was analyzed by flow cytometry (b-d). MFI = mean fluorescent intensity. NS = not significant, *** P < 0.001. One-way ANOVA and Dunnett’s multiple comparison test