Fig 2: VWF is transported in circulating EVs from GBM patients. (A) Equal volume of protein lysates from EV fractions, obtained with SEC followed by ultracentrifugation, and their corresponding plasma, from one GBM patient and one healthy donor, were separated by SDS-PAGE and analyzed by immunoblot for VWF, FCN3, and HSP70. Blots are representative of n = 3. (B) mRNA expression of von Willebrand factor (VWF) and ficolin-3 (FCN3) in healthy subjects and GBM patients estimated from The Cancer Genome Atlas (TCGA, n > 500 for GBM patients). (C) Anomalies currently reported in VWF and FCN3 genes. (D) Equal volume of protein lysates from EV fractions (3 GBM and 3 Healthy) were separated by SDS-PAGE and analyzed by immunoblot for VWF, GM130 (putative intracellular membrane protein contaminant), and Apolipoprotein A1 (APOA1, plasmatic protein). Healthy donor plasma and total cell lysate from GBM cells serve as controls. Blots are representative of n > 7 individual samples. (E) Densitometric analysis was performed on 7 and 8 independent EV preparations from Healthy and GBM liquid biopsies, respectively. (F) Concentration of VWF in plasmatic EVs from healthy donors and GBM patients (n = 20). Mann–Whitney test, *p < 0.05, **p < 0.01, ***p < 0.001. S.D. are shown in panel E and S.E.M. in panel F.

Fig 3: EC differentiation of hiPSCs from patients with ESRD and HCs.(A) Schematic overview of EC differentiation. (B) Expression levels of the EC markers CD31, CD34, CD133, VWF, Flk-1, and Flt in purified hiPSC-ECs before and after CD31+ isolation, as determined by immunofluorescence staining with quantitative analysis. Scale bar represents 50 µM. **p < 0.05 relative to HC-iPSC-ECs. (C) CD31+ expression on hiPSC-ECs differentiated from patients with ESRD and HCs, as determined by flow cytometry analysis. **p < 0.01 relative to HC-iPSC-ECs.EC, endothelial cell; ESRD, end-stage renal disease; Flk-1, fetal liver kinase 1; Flt-1, vascular endothelial growth factor receptor 1; HC, healthy control; hiPSC, human induced pluripotent stem cell; SSC, side scatter; vWF, von Willebrand factor.