Fig. 2

Dose-dependent response of 2D iBMEC monolayers to chronic and acute oxidative stress. a Heatmap of iBMEC TEER during chronic exposure to H₂O₂ at various concentrations and corresponding controls (no addition, or vehicle addition). Continuous 1 mM H₂O₂ exposure resulted in a gradual loss of barrier function over 10 days that models chronic oxidative stress. Each cell represents the mean of three technical replicates across n = 3 biological replicates in all conditions. b Heatmap of iBMEC TEER during acute exposure to H₂O₂ at various concentrations and exposure times and corresponding controls (no addition, or vehicle addition). 10 mM H₂O₂ exposure for one hour resulted in significant but recoverable loss of barrier function over 10 days that is consistent with acute oxidative stress. Each cell represents the mean of three technical replicates across n = 7 biological replicates in all conditions. c Representative images from DAPI and ROS assay staining showing the negative control with minimal ROS accumulation, continuous 1 mM H₂O₂ exposure with minimal ROS accumulation, and acute 10 mM H₂O₂ exposure with visible ROS accumulation and cell loss. Yellow dotted line indicates a region of cell loss (no nuclei). d, e Quantification of cell counts over time following chronic and acute exposure, respectively. f, g Quantification of intracellular ROS per cell over time following chronic and acute exposure, respectively. Statistical tests were conducted versus control at each timepoint. n = 3 biological replicates for all conditions and time courses for (d–g)