Figure 5.

Cyclic nucleotides do not affect platelet spreading, P‐selectin exposure and Ca²⁺ elevation on HLECs. Washed platelets (5 × 10⁸ per mL) were treated with 1 μm PGI₂, 10 μm forskolin, 1 mm GSNO or vehicle and allowed to spread on a HLEC monolayer for 1 h at 37 °C. Slides were stained for podoplanin, CD41 and P‐selectin and imaged using confocal microscopy (scale bar = 20 μm) (A). Area of attached platelets (B) and P‐selectin/CD41 intensity (C) were evaluated (n = 3). 2 × 10⁷per mL Fura‐2‐loaded platelets were treated as above or with 5 μm PRT060318 (PRT) and allowed to attach on a HLEC monolayer. Attachment of platelets was recorded for 20 min. Average [Ca²⁺]_i concentrations were calculated and reported as mean ± SEM (n = 3). Statistical differences were evaluated with anova and Dunnet's post‐test (*P < 0.05) (D).