Figure 5.

ADP-induced platelet aggregation. Washed human platelets were incubated with 2 µM CsA, 20 µM bongkrekic acid (BK) or 10 µM FK-506 (FK), or their respective vehicles (1% ethanol for CsA, 2 mM NH₄OH for bongkrekic acid and 0.5% DMSO for FK-506) for 30 min at 37 °C. The conditioned human and the washed murine (wild-type: WT or CypD^−/−: KO) platelets were supplemented with 6 µM fibrinogen to support aggregation right before it was initiated by the addition of 10–50 µM ADP (to give a maximal aggregation value of at least 30%) and 2.5 mM CaCl₂. PBS was used as background for 100% transparency, and aggregation was recorded for 30 min. Aggregation curves from representative experiments using human (a) or murine (b) platelets are shown. (c,d) The measured initial slope and maximal aggregation values are shown in relative units (the mean value of the replicates of the vehicle control in each independent set of measurement was considered to be 1). ‘No additive’ designates samples incubated with the respective vehiculum of the inhibitors. Data from duplicates from at least three independent experiments and SE are shown. Asterisks mark differences significant at P < 0.05 between the indicated pairs of samples according to the Kolmogorov-Smirnov test. *a: P = 0.0438, *b: P = 0.0111, *c: P = 0.0026, *d: P = 0.0361, *e: P = 0.0111, *f: P = 0.0275 *g: P = 0.0042.