Nrf2 critically regulates BBB endothelial structural integrity and function.

<p><b>(A)</b> Nrf2 knock-down efficiency by gene silencing as determined by IF or western blot analyses of Nrf2 (red) and its downstream target, NQO1, in hCMEC/D3 cells transfected with Nrf2-specific or scramble siRNA (n = 3/condition). <b>(B)</b> Effects of Nrf2 silencing on endothelial tight and adherence junction protein expression in hCMEC/D3 monolayers. Respective bands with β-actin as loading control were shown at the bottom of the graphs for each protein (n = 3/condition). <b>(C)</b> Effects of Nrf2 knock-down on TEER and paracellular permeability to the mixture of labeled dextrans (4 and 10kDa) across confluent hCMEC/D3 monolayers cultured on 12 or 24-well Transwell inserts and transfected with scramble or Nrf2 targeting siRNA (n = 5-6/condition). Images were captured at 40X (scale: 100 μm) and merged with DAPI. Data were expressed as mean ± SEM (% of euglycemic scramble control). ***<i>P</i> < 0.01, **<i>P</i> < 0.01, *<i>P</i> < 0.05, vs. control.