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. 2022 Jan 25;7(4):e153560. doi: 10.1172/jci.insight.153560

Figure 1. Characterization of airway EVs following i.t. treatment with LPS.

Figure 1

(A) EVs were harvested from BALF 24 hours following saline or LPS (35 μg) treatment of 11-week-old A/J mice (n = 10 per group). (B) Flow cytometric analysis of the cellular composition of BALF following LPS or saline treatment. (C) Size distribution of airway EVs (LPS and saline treated) determined using NanoSight measurements following purification via differential ultracentrifugation. (D) Quantification of surface NE on airway EVs of LPS- and saline-treated mice by bead-based flow cytometric analysis. (E) Analysis of NE activity of airway EVs (LPS and saline) using an NE-specific FRET assay. (F) NE activity for EVs from LPS-treated Elane–/– mice or WT mice treated with α-1AT (1 μg) or depleted of Ly6G+ EVs. (G) NE activity for LPS EVs from E pretreated with NE Inhibitor II (NE INH. II; 20 μM) prior to the assay. Data are shown as median and IQR (n = 3 replicates per experiment). Statistical analyses were performed using Wilcoxon’s signed-rank test; ****P < 0.0001.