Confocal imaging of the tight junction proteins cotreated with VEGFA and DMSO, SB431542, or RepSox. The immunocytochemistry images for each tight junction proteins with cotreatment of compounds and VEGFA in (A) hPSC-ECs and (B) hBMECs. The proteins Claudin-5, ZO-1, and Occludin in red channel, and VE-Cadherin in green, and DAPI in blue. (C) The respective quantification of the fluorescence intensity of the individual proteins (Claudin-5, ZO-1, and Occludin) in the area of interest where Claudin-5, ZO-1, or Occludin signal colocalized with VE-Cadherin as the area of interest. Images shown are representative image set with magnification at 20×. Columns are means ± SD. (Scale bars: 50 μm.) The imaging was performed with at least three replicates. *P < 0.05, **P < 0.01, ***P < 0.001.