Fig. 3

Robo4 silencing degrades the expression and organization of TJ molecules after γ-radiation exposure. A–E Western blotting analysis of claudin-5, occludin, PECAM-1, and ZO-1 normalized with endogenous beta-actin protein. The relative protein expression were quantified using Image J software. F Immunostaining for claudin-5 (red) and DAPI staining (blue) in irradiated Robo4 knockdown and control microvascular EC samples. G Flow cytometric analysis of PECAM-1 expression levels after Robo4 knockdown and irradiation. H Confocal imaging of ZO-1 (red) and DAPI (blue) in Robo4 silenced and controlled ECs with or without gamma radiation exposure. Scale bars = 20 µm. For all experiments, n ≥ 3, and error bars represent std. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001