Fig. 3

Endothelial cell migration and ECM production is enhanced by LP. A The effect of LP treatment on the migration potential of HUVEC was analyzed via wound migration assay. Confluent cells were wounded using a p1000 pipet tip and were either left untreated or treated with LP for 24 h. A wound migration assay showed that LP treatment for 30 s and 60 s promoted endothelial cell migration. The mean denuded zone was determined by calculating the ratio of the average denuded zone area to that of the control zone. The asterisks indicate statistically significant differences. ***P < 0.001. B LP regulated the total protein expression of VE-cadherin and ECM (p-FAK(Y397), FAK, p-Src(Y418), and Src) proteins, as shown by western blotting. HUVECs were treated with LP for 30 s and 60 s and then cultured for 24 h. α-tubulin was used as the loading control. Band intensities were measured and are represented as graphs. *P < 0.05, **P < 0.01. C A representative zymogram demonstrates that increasing concentrations of LP were associated with a selective increase in MMP-2 activity. Band quantitation performed to examine the effects of the activity of 72 kDa and 62 kDa MMP (expressed as the ratio of the control). **P < 0.01, ***P < 0.001. D Relative expression levels of MMP-2 and MMP-9 mRNA were determined using real-time PCR. No significant changes were observed in MMP-9 activity. F Immunocytochemical assays for VE-cadherin and p-FAK. E VE cadherin was decreased in LP-treated cells; scale bar = 20 μm. F Focal accumulation of FAK was significantly increased in LP-treated cells. The areas outlined in red are enlarged in the side panels; scale bar = 30 μm