Fig. 1

Human BMEC-like cells internalize EEVs. (A-B) Representative confocal images of control BMEC-like cells stained for ZO-1 (white), DAPI nuclear stain (A), and time-course quantification of EEV internalization (CellMask Deep red, white arrows) (A-B) from 10 min to 48 h incubation times. Scale bars: 20 μm. (C) Western blot-based quantification of tight junction protein levels for VE-cadherin, ZO-1, and Claudin 5 in BMEC-like cell monolayers after a 48 h incubation with EEVs. Protein levels are normalized to actin loading control. Statistical analysis: Error bars represent mean + SEM. Statistical analysis was performed using two-way ANOVA followed by Bonferroni’s multiple comparisons test. In (B), two different iPSC lines (line IDs 38554 and 41658) and in (C), three different iPSC lines (line IDs 38554, 41658, and 36091) were treated with EEVs produced from erythrocytes of two different healthy donors (age = 30 years old). Abbreviations: Ctrl, control; DAPI, 4′,6-diamidino-2-phenylindole; EEV, extracellular vesicles derived from erythrocytes; h, hours; min, minutes; ns, not significant; ZO-1, Zonula occludens-1