Fig 2: Biological impact of blocking IGFBP-2 in clot formation and endothelial cell function. Changes in hemostatic parameters after IGFBP-2 blockade in thromboelastometry assays with blood samples of healthy subjects (n = 5) (A–C). CFT: Clot formation time, MCF: maximum clot firmness, LOT: lysis onset time. * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. ctrl (n = 5/group). Number of transmigrated THP-1 monocytes in presence or absence of a neutralizing antibody against IGFBP-2 across TeloHAECs (n = 5/group) (D). Vascular permeability to BSA over time in hCMEC/D3 control cells (black solid line), or thrombin stimulated cells with (dotted line) or without (dashed line) a pretreatment with a neutralizing IGFBP-2 antibody (E). Four independent experiments were performed, with two technical replicates in each one. * p < 0.05 vs. thrombin at 1 h.

Fig 3: Gene expression and western blot of selected candidates in plasma EVs from IS patients. (A–C) mRNA levels, presented as Ct values, of IGFBP-2, NECTIN-2, and PECAM-1 assessed using RT-qPCR in plasma EVs from n = 10 IS patients (AT = 3: black dots, CE = 4: grey dots, and ESUS = 3: white dots). Undetected transcripts were assigned Ct = 40. (D–F) Western blot of plasma EVs from IS patients (n = 3 AT and 3 CE) showing selected candidates and their expected molecular weights (MW). EVs 1 to 3 are from AT patients, and EVs 4 to 6 are from CE patients.

Fig 4: Circulating levels of IGFBP-2, NECTIN-2, and PECAM-1 according to patient clinical etiology. IGFBP-2 and NECTIN-2 concentrations were increased in CE and ESUS patients compared to AT (A,B). No differences were observed in blood levels of PECAM-1 according to IS origin (C). (n = 19–23 AT, n = 40–54 CE and n = 19–23 ESUS). * p < 0.05 vs. AT.