Fig. 5

Response of three-dimensional iBMEC microvessels to physical injury. A Representative photographs of Evans Blue leakage induced by laser ablation of an area corresponding to 5–10 cells. B Live-cell imaging of TJ-labeled iBMEC microvessel recovery following laser ablation. C Time course of wound size (n = 4 microvessels). D Centroid tracking of cell neighbors surrounding a wound during healing (red dots), as well as cell neighbors of an apoptotic cell (black dots). Cell neighbors of ablation display directed migration towards the centroid of the wound. E Cell size distribution immediately after ablation and following wound closure of a total monolayer area of 35,000 µm² (n = 353 cells in open wound and n = 290 cells in closed wound). F Heatmap comparing changes in cell morphology of nearest neighbors surrounding a wound during healing, relative to quiescent cells and neighbors of an apoptotic cell. 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. S11–S13; all analysis conducted on TJ-labeled iBMECs