Fig 2: Identification of Angpt1 in PCEVs by proteomic analysis and ELISA. a, b Gene ontology (GO) results of differentially expressed genes (DEGs) from the GSE195917 and GSE190809 datasets categorized into biological process (BP), cellular component (CC), and molecular function (MF). c, d Heatmap in vascular endothelial function pathways. e, f GO enrichment analysis of upregulated DEGs in PCEVs and heatmap in vascular endothelial function pathways. g Expression levels of Angpt1 in PCs, SMCs, PCEVs, and SMCEVs as determined by ELISA (n = 3). h Quantification by multiplex sandwich ELISA of Angpt1, VEGF, and TGF-β1 concentrations in PCs and PCEVs (n = 3). ***P < 0.001

Fig 3: Angpt1 carried by PCEVs regulates VEC migration, proliferation, and tube formation. a Representative microphotographs of the scratch migration assay and the scratched area were shown (n = 8). Scale bars: 100 μm. b CCK8 proliferation assay of VECs (n = 8). c Tubular structures were photographed, and the number of nodes and junctions were calculated using the ImageJ software (n = 8). Scale bars: 50 μm. d Western blot assay was performed to detect Angpt1, p-Tie2, Tie2, p-PI3K, PI3K, p-Akt, Akt, p-NF-κB, NF-κB, and β-actin protein levels (n = 3). ***P < 0.001 vs Nor, ##P < 0.01 vs. LPS, @P < 0.05, @@P < 0.01 vs. PCEV (one-way ANOVA)

Fig 4: Effects of Angpt1 carried by PCEVs on VEC barrier functions following LPS administration in vitro. a Illustrative images of the internalization of PKH-26-labeled PCEVs in VECs. Scale bars: 10 μm. b PCEVs were added to VECs, and the TER of each group was measured (n = 3). c PCEVs were added to VECs, and FITC-BSA penetration of each group was measured (n = 8). d VECs treated with PCEVs were analyzed by western blotting (n = 3). e VECs treated with PCEVs were analyzed by immunofluorescence for ZO-1 and VE-cadherin. Scale bars: 10 μm. ***P < 0.001 vs. Nor, ##P < 0.01 vs. LPS, @@P < 0.01 vs. PCEV (one-way ANOVA)