Adenovirus-mediated KLF11 overexpression in mouse BMEC cultures preserves the integrity of tight junction proteins after OGD. Cultured mBMECs were infected with Ad. LacZ or Ad. KLF11 for 48 h prior to 16 h or 4 h OGD followed by 24 h or 48 h reperfusion. (A-B) The mRNA expression levels of ZO-1 and occludin were determined by qPCR and normalized to cyclophilin (n=3 per group). (C-D) Expression levels of tight junction proteins, ZO-1 and occludin were evaluated by western blotting. β-Actin was used as an internal loading control. Data represent three independent experiments and representative blots are displayed. n=3 per group. * P < 0.05 vs. Ad. LacZ + non-OGD group, # P < 0.05 vs. Ad. LacZ + OGD group. (E) Representative images and quantification of ZO-1 (green) and occludin (green) in cultured mBMECs after 4 h OGD, followed by 24 h reoxygenation. (Scale bar: 25 μm.) Adenoviral KLF11 overexpression suppressed OGD-induced disruption of junctional proteins, ZO-1, and occludin. n=3 per group. * P < 0.05 vs. Ad. LacZ + non-OGD group, # P < 0.05 vs. Ad. LacZ + OGD group. H16R0 = 16h OGD + 0h reperfusion, H16R24 = 16h OGD + 24h reperfusion, H16R48 = 16h OGD + 48h reperfusion, H4R0 = 4h OGD + 0h reperfusion, H4R24 = 4h OGD + 24h reperfusion.