Fig. 7.

Effects of VEGF on the paracellular permeability of b.End3 cells. Cells were treated with 100 μM CoCl₂, HIF-1α inhibitors (YC-1, 2ME2), HIF-1α-specific siRNA (siRNA) and negative control siRNA (NC) under the same conditions as described in Fig. 6. VEGF antibody (Ab-VEGF) at 100 ng/ml (final concentration) was added to the cell culture medium 2 h before the high glucose (30 mM) treatment. Cells were incubated in high glucose medium with or without Ab-VEGF for 6 days. The cell culture medium was changed every 2 days with addition of the inhibitors or VEGF antibody to the new culture medium. Cells treated with 30 mM l-glucose (30 L) in the presence of 5.5 mM d-glucose were used as an osmotic control. a Representative Western blots of VEGF expression in b.End3 cells with β-actin as a protein loading control. b Summary of VEGF protein expressions from three independent experiments. Values were normalized to β-actin and control (5.5 mM glucose). c Effect of Ab-VEGF on permeability of cells exposed to 30 mM high glucose treatment for 6 days. d Immunostaining ZO-1 and occludin in b.End3 cells exposed to 30 mM glucose for 6 days. Scale bar 22 μm. Data are means ± SEM, n = 3. *p < 0.05 vesus control (5.5 mM glucose), ^# p < 0.05 versus 30 mM glucose