Fig. 5.

DV-induced NETs contribute to changes in vascular permeability. a and b Neutrophils were incubated with DV, EVs (from 8 × 10⁶ platelets, including both MVs and EXOs), or DV-EVs (from 8 × 10⁶ DV-activated platelets) and overlaid on HMEC-1 monolayers in presence or absence of DNase I (100 U/ml) at 37 °C for 3 h (a). Alternatively, neutrophils were pretreated with isotype control, anti-CLEC5A mAb, and anti-TLR2 mAb at 37 °C for 1 h, followed by incubation with DV or EVs (b). These assays used 1.5 × 10⁸ EVs per experiment (a and b). Endothelial monolayer permeability was measured by determining HRP activity in media from the lower chamber. c Stat1^−/− mice were challenged by intravenous administration of DV, in the presence or absence of DNase I, followed by injecting Evans blue (i.v.) at 1 h before sacrifice on day 5 post-infection (n = 4 in saline (vehicle) group, n = 3 in DNase I group). d Stat1^−/− (n = 6 in saline and isotype control group, n = 4 in anti-TLR2 mAb group) and stat1^−/−clec5a^−/− (n = 5 in saline group, n = 4 in isotype control and anti-TLR2 mAb group) mice were pre-treated with anti-TLR2 mAb or isotype control, followed by inoculation with DV (NGC-N) via intraperitoneal route. Evans blue was injected intravenously at day 5 post-infection, 1 h before mice were sacrificed. Plasma leakage of visceral organs was determined by measuring absorbancies at 610 nm for each organ. Data are mean ± SEM from at least three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 (Student’s t-test). Source data are provided as Source Data file