Fig. 1

IR decreases the expression of EC junctional molecules but upregulates Robo4 expression. A–C RT-qPCR shows the relative mRNA expression of the TJs, GJs, and AJs. Gene expression results were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and beta-actin endogenous control and represented as fold change in gene expression. The error bars stand for the standard deviation from three separate independent experiments. D and E Detection of TJs (claudin-5 and occludin) protein expression by immunocytochemistry and western blot, respectively. F and G Analysis of gap junction (connexin 43) protein expression using immunofluorescence and immunoblotting. H and I The distribution and expression of VE-cadherin analysis with immunofluorescence, cycloheximide chase experiment, and western blot. J and K mRNA and protein detection of Robo4 in microvascular ECs after irradiation using RT-qPCR and WB. M Effect of irradiation on cell survival and proliferation of ECs. For all experiments, scale bars = 20 µm. n ≥ 3, and error bars represent std. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001