Fig. 1.

Gating strategy for flow cytometric characterisation of MVs. a) A 100-1000 nm MV size gate was established based on 200 nm and 900 nm polystyrene beads (MegaMix) and transferred to all samples. b) Next, a gate was set on FITC-H at the 99th percentile of unlabelled samples. c) MVs were defined as PS+ events based on binding of lactadherin-FITC. e & f) The double negative population was defined based on density (magenta gate), and bi-variate gates were placed at the 99th percentile of the double negative population. Finally, the gates were applied to all PS+ events in the corresponding sample to define MMVs (e), EMVs (f), and the expression of CD36 on these phenotypes (e & f). d) TruCount® beads (magenta gate) were quantified and used to calculate absolute concentrations of MVs. g) Gating hierarchy utilised for defining the MV phenotypes in the current study. PS: Phosphatidylserine; PFP: Platelet-free plasma; MMV: Monocyte microvesicles; EMV: Endothelial microvesicles