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. 2015 Sep 18;5:14090. doi: 10.1038/srep14090

Figure 3. Influences of Src signaling on endothelial permeability.

Figure 3

(a) PP2 prevented AGE-induced hyperpermeability. HUVECs were treated with PP2 90 min before AGEs (100 μg/mL, 8 h) application. Culture medium was used as control. n = 3 *P < 0.05 versus control, #P < 0.05 versus AGEs. (b) Src siRNA caused reduction in Src protein level compared with control siRNA. HUVECs were transfected with Src siRNA and control siRNA for 48 h, after which cells were lysed and detected for Src protein level by WB. (c) Src siRNA prevented AGE-induced hyper-permeability. HUVECs were transfected with siRNA for 48 h before exposure to AGEs (100 μg/mL) for 8 h. n = 3, *P < 0.05 versus control, #P < 0.05 versus control siRNA with AGEs. (d,f) The effects of pcDNA3/flag-Src (WT-Src), K298M and Y530F on expression of Src were detected by western blotting. (e,g,h) The effects of WT-Src, K298M and Y530F on EC permeability. Cells were transfected with WT-Src, K298M and Y530F with or without stimulation of 100 μg/mL AGEs for 8 h. TER and Pd were detected. n = 3, *P < 0.05 versus control or mock, #P < 0.05 versus AGEs or mock with AGEs. (i) The effects of Src on the distribution of F-actin induced by AGEs. ECs were treated with or without 100 μg/mL AGEs for 8 h, followed by rhodamine-phalloidin staining, and F-actin was revealed by a laser confocal microscopy.