Fig. 4

Phenotypic characterization of migratory capacity and barrier permeability in PCV endothelial cells. a Gene set enrichment analysis against Gene Ontology (Biological processes) database reveals enriched processes from PCV upregulated genes shown here ranked by Bonferroni corrected p values. Right panel shows violin plots that represent calculated module scores for the processes indicated. b Wound healing assays carried out to assess migratory capacity of BOECs 21 h post-scratch after 24 h of static or rotated (heterogeneous flow) culture. Left panel shows representative images from heterogeneous flow condition with yellow outlines indicating cell-free wound regions after 21 h, scale bar, 2000 μm. Box and whiskers plots showing median with minimum and maximum. n indicates number of cell lines evaluated for each group while p value is from two-tailed t-test, ** p < 0.01. c Heatmap showing average expression of cells from each individual BOEC sample in the indicated cell states. d Violin plots of glycocalyx-related genes found to be significantly differentially expressed between PCV and normal BOECs, *adj. p < 0.01. e In vitro vascular permeability imaging assay using biotinylated-gelatin-coated surfaces. Intercellular gaps were revealed by Neutravidin-FITC staining after 24 h of static or rotated (heterogeneous flow) culture. Shown are representative frames from different BOEC lines, scale bar, 200 μm. Box and whiskers plots showing median with minimum and maximum. Area was normalized against average FITC area of normal BOEC lines. n indicates number of cell lines evaluated for each group while p value is from two-tailed t-test, *** p < 0.001, **** p < 0.0001