Figure 5. We used Boyden chambers and paxillin immunofluorescence staining assays to determine that effects of heparanase expression levels on human retinal vascular endothelial cell migration. A: The human retinal vascular endothelial cells (HRECs) that are stained with crystal violet are those that migrated across the extracellular matrix (ECM) in the specialized migration chamber by migrating through the polycarbonate membrane to the lower surface of the membrane (original magnification 200×). B: Paxillin immunofluorescence staining indicated positive paxillin protein expression in every group. HPSE-LV significantly inhibited HREC paxillin protein expression compared with the Con-LV-Glu30 and Glu30 groups. The paxillin protein expression in the Glu30 group was significantly higher than in the HPSE-LV-Glu30 and Glu5 groups (the yellow arrow indicates paxillin). C: Heparanase small interfering RNA recombinant lentiviral vector (HPSE-LV) significantly decreased the HRECs’ migration ability. Bars indicate the mean±SD (p<0.05). The HPSE-LV-Glu30 group showed a markedly decreased migration ability compared to the Con-LV-Glu30 (p<0.01) and Glu30 groups, which migrated significantly more than the HPSE-LV-Glu30 and Glu5 groups (●p<0.01 versus Glu5; ▲p<0.01 versus HPSE-LV-Glu30). Each group experiment was repeated 3 times (n=3).