Gran-MVs are the MV subset that mediates the thrombolytic activity. (A) Flow cytometric monitoring of MVs after selective depletion of MV subsets: platelet-derived MVs (P-MVs), erythrocyte-derived MVs (ery-MVs), and granulocyte-derived MVs (Gran-MVs). Immunomagnetic separation was performed using beads coated either with CD41 (red circle), CD235a (red square), or CD15 (red rhombus). Negative control experiments were performed in parallel using beads coated with an irrelevant antibody (IgG, blue symbols); n = 3. (B) The MV-PGC test was performed on MVs after the selective depletion of CD41⁺, CD235a⁺, or CD15⁺ MV subpopulations or after a control IMS (IgG); n = 3, CD41: red circle; CD235a: red square; CD15: red rhombus; IgG: blue circle. (C) The thrombolytic effect of MVs depleted of CD15-MVs or those subjected to a control IMS depletion (IgG) was evaluated in the lysis front retraction model at 0, 24h, and 48 hours in the presence of 10⁷ MVs. (D) The lysis area was calculated at 48 hours, considering the entire thrombus surface (4.3 mm²), using ImageJ software. *P < .05; **P < .01. (E-F) Thrombolysis mediated by Gran-MVs (5 × 10⁶ MVs) in a lysis front retraction model. Lysis is amplified by the saturation of uPAR with exogenous uPA (uPA-Gran-MVs) on the MV surface. Representative fluorescence microscopy images (E) display the retraction of the lysis front (white arrows) at 0, 6, 24h, and 48 hours; black bar = 1000 µm. The lysis area (F) was calculated considering the whole thrombus surface (150 mm²) using ImageJ software; uPA-Gran-MV: red circle; Gran-MV: blue circle; SPN: green circle; n = 5. *P < .05; **P<.01