Figure 4.

cAMP‐elevation marginally affects CLEC‐2‐dependent spreading. 2 × 10⁷ per mL washed platelets were treated with PGI₂ (1 μm), forskolin (10 μm) or GSNO (1 mm) before spreading on coverslips coated with 10 μg mL⁻¹ recombinant podoplanin or CLEC‐2 mAb. Control experiments with thrombin‐activated platelets (0.1 U mL⁻¹) on 100 μg mL⁻¹ fibrinogen were performed. Coverslips were imaged using differential interference contrast (DIC) microscopy (A). Platelet areas are expressed as mean of mean area/experiment ± SEM. One hundred platelets/experiment were analyzed. Statistical differences were evaluated by anova and Dunnet's post‐test (**P < 0.01) (B). Results are expressed as mean of mean number of platelets/experiment ± SEM (n = 3–6) (C).