Fig. 4. HMB-001 does not influence key platelet properties.
a,b, Whole blood from healthy donors (n = 3) was incubated with 0 or 100 nM HMB-001 and buffer or platelet agonists (PAR-1 AP, PAR-4 AP, ADP, CRP-XL or U-46619) for 10 min. Platelet P-selectin expression (a) and fibrinogen binding (b) were measured with FACS. Data are expressed as mean MFI ± s.d. c, PRP from healthy donors (n = 3) was incubated with 0 or 100 nM HMB-001 and activated with ADP, epinephrine, collagen or PAR-1 AP with stirring at 900 r.p.m. Platelet aggregation was monitored with light transmission for 15 min. Data are expressed as the maximal amplitude of the aggregation trace and represent mean ± s.d. d, TLT-1 shedding. Washed platelets from healthy human donors (n = 3) were stimulated with collagen in an aggregometer for 0–60 min. Platelet fractions and supernatant were separated with centrifugation and subjected to SDS–PAGE. TLT-1 was visualized with western blotting. The intensity of the 17-kDa band was analyzed with Empiria Studio 2.1 software. Data represent mean ± s.d.