Fig. 3.

Time-dependent effects of high glucose on the expression and arrangement of ZO-1, occludin and claudin-5 in b.End3 cells. Cells were treated with glucose at 5.5, 30 and 50 mM for 1, 3, 6 or 10 days. l-Glucose at 50 mM in the presence of 5.5 mM d-glucose was used as osmotic control (50 L). a Representative Western blots of ZO-1, occludin and claudin-5 after high glucose treatments. β-actin served as a protein loading control. b Summary of the protein level of ZO-1. c Summary of the protein level of occludin. d Summary of the protein level of claudin-5. e Representative images of ZO-1, occludin and claudin-5 immunostaining after cells were exposed to 30 mM glucose for 6 days. Scale bar 22 μm. l-Glucose at 50 mM in the presence of 5.5 mM d-Glucose was used as osmotic control. Values in b, c and d were normalized to β-actin and control (5.5 mM glucose). Values are means ± SEM, n = 3. *p < 0.05 versus 3 day control (5.5 mM glucose), ^# p < 0.05 versus 6 day control (5.5 mM glucose), ^@ p < 0.05 versus 10 day control (5.5 mM glucose)