Fig. 1: Flow cytometry of platelets and platelet-derived microvesicles.

Platelets were incubated for 15 min (light bars) and 60 min (dark bars) under the following conditions: untreated platelets (negative control), platelets treated with the KKO/PF4 and RTO/PF4 complexes, KKO, RTO, and PF4 alone as well as Ca²⁺-ionophore A23187 (positive control). Final concentrations: 10 µg/ml PF4, 50 µg/ml KKO, and RTO, 10 µM Ca²⁺-ionophore. a Annexin V-positive platelets as a fraction of all gated platelets taken as 100%: untreated platelets (n = 17/13 for 15 min/60 min, respectively), KKO/PF4 (n = 16/13), RTO/PF4 (n = 3/3), KKO (n = 16/13), RTO (n = 3/3), PF4 (n = 16/13), and A23187 (n = 14/12). b CD62P-positive platelets as a fraction of all gated platelets taken as 100%: untreated platelets (n = 8/7), KKO/PF4 (n = 8/7), RTO/PF4 (n = 3/3), KKO (n = 8/7), RTO (n = 3/3), PF4 (n = 8/7), and A23187 (n = 8/7). c Number of CD41- and Annexin V-double-positive microvesicles normalized by the total number of gated platelets in the corresponding sample: untreated platelets (n = 8/9), KKO/PF4 (n = 8/9), RTO/PF4 (n = 3/4), KKO (n = 8/9), RTO (n = 3/4), PF4 (n = 8/9), and A23187 (n = 8/9). “n” is the number of experiments with platelets from independent donors. *P < 0.05 compared to a corresponding negative control. The differences between 15 and 60 min are insignificant