Effects of compound 8 (10 μM) on the barrier integrity of human brain microvascular endothelial cells (hCMEC/D3) in the absence and presence of oxidative stress promoted by tert-butyl hydroperoxide (tBHP, 350 μM).

All treatments were performed for 4 hours. A: Transendothelial electrical resistance (TEER) measurement. B: Permeability measurement for the paracellular marker molecule FITC-dextran 4 kDa (FD4). C: Permeability measurement for the transcellular marker molecule Evans blue labeled-albumin (EBA, 67 kDa). Data are presented as mean ± standard deviation (SD; n = 4). Data analysis was performed using one-way analysis of variance (ANOVA) followed by Bonferroni’s multiple comparisons test. Statistical significance: ***p < 0.001, compared to the control group, and ^###p < 0.001, compared to the tBHP group. D: Immunocytochemistry for β-catenin and zonula occludens-1 (ZO-1) junctional associated molecules. Green: junctional staining. Blue: cell nuclei. Bar: 20 μm.