Fig. 5

Circulating EVs from newborn rats with VILI are selectively loaded with caspase-1 and can cross the blood brain barrier. Nanoparticle tracking analysis of EVs isolated from controls (A), low Vt (B), and high Vt (C) show a distinct peak between around 100–120 nm, which corresponds to the exosome particle size. These EVs were enriched for the exosome markers CD9 and CD81 (D). Adoptive transfer by tail vein injection of fluorescence-labeled EVs shows that these EVs can cross the blood brain barrier and localize to the brain on in vivo and ex vivo imaging (E). Western blot of EVs extracted from controls and ventilated animals shows increased activated caspase-1 (F) but not GSDMD (G) in the EVs from animals with ventilation induced lung injury. Data are presented as box plots showing the median, 25th to 75th percentile, and minimal to maximal value normalized to control. N = 4 animals/group for nanoparticle tracking analysis. N = 10 controls, 6 low Vt, and 8 high Vt for EV Western blot analysis. Data was analyzed by ANOVA with Dunnett’s post-test for multiple comparison, p values < 0.05 are shown on bars