Fig. 6

Response of three-dimensional iBMEC microvessels to chemical injury. A, B Real-time imaging of iBMEC microvessels in response to perfusion with menadione or melittin. A Red arrows indicate delamination of the endothelium and white arrows denote sites of leakage of 10 kDa dextran. B Red arrows indicate cell collapse and white arrows denote sites of leakage of 10 kDa dextran. C, D Comparison of 10 kDa dextran permeability and cell turnover between control, menadione-exposed, and melittin-exposed iBMEC microvessels (n = 4 or more). Significance analysis performed using a Kruskal–Wallis test and post hoc Dunn’s multiple comparisons test relative to control. E Cell size distribution before and after melittin exposure (n = 581 cells pre-melittin and 329 cells post-melittin). F Heatmap comparing changes in cell morphology for apoptotic cells and cell loss induced by melittin exposure, as well as their respective nearest neighbors (NN). Only squares corresponding to a statistically significant change are shaded by magnitude of change (log₂FC) and direction of change (blue: increasing over time, red: decreasing over time) labeled with asterisks representing significance as calculated using the maximally significant result between a Wilcoxon matched-pairs signed rank test to compare first and last time points and using an F test to determine if linear regression of the morphological time course displayed a statistically non-zero slope. Data are presented as mean ± SD. *p < 0.05, **p < 0.01, and ***p < 0.001. See also Additional file 1: Figs. S12, S13; all analyses conducted on TJ-labeled iBMECs