Figure 3 from HuR-Regulated Extracellular Vesicles Promote Endothelial Cell Remodeling in Pancreatic Cancer

WT EVs directly alter endothelial cell transcriptome, migration, barrier function, and tube formation. A, HPaMECs were treated with WT vs. HuR-KO EVs for 24 hours before RNA-seq and phenotypic assessment of migration, monolayer permeability, and tube formation. B, Volcano plot of differentially expressed genes in HPaMECs treated with PANC-1 HuR WT (blue, left) vs. HuR-KO (orange, right) EVs for 24 hours (n = 3). C, Gene Ontology biological process analysis of the top seven pathways enriched in WT EV–treated and the top five pathways enriched in HuR-KO EV–treated HPaMECs (n = 3). D, Transwell migration of HPaMECs treated with PANC-1 WT vs. HuR-KO EVs over 24 hours and quantified with crystal violet staining (n = 3). Functional analysis of HPaMECs treated with media alone, PANC-1 WT, or HuR-KO EVs for 24 hours and monitored for (E) monolayer permeability quantified by dextran movement across the endothelial cell monolayer and (F) representative phase-contrast tube formation images with full field of view and region of interest (scale bar, 250 µm). G, Tube formation quantification for total tube length (px), total branching points, and total loops (n = 3). P values were calculated using an unpaired two-tailed Student t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; Card. M., cardiac muscle; CNS, central nervous system; Contr., contraction; EC, endothelial cell; Kid., kidney; Lym., lymphatic; Morphog., morphogenesis; ns, not significant; PCP, planar cell polarity; Resp., respiratory; VEGF, vascular endothelial cell growth factor.