Journal: Molecular Neurobiology

Article Title: Endothelial Neurogranin Regulates Blood–Brain Barrier Permeability via Modulation of the AKT Pathway

doi: 10.1007/s12035-024-04522-9

Figure Lengend Snippet: Decreased Ng expression increases permeability in hCMEC/D3 cells. A Methodology for the TEER assay. Continuous TEER was measured over a monolayer of hCMEC/D3 cells cultured on transwell filters. The line graph shows the development of TEER of lipofectamine control (Mock) cells versus cells treated with Ng-siRNA, indicating a sustained decrease in TEER in Ng-siRNA treated cells. # p < 0.05; 2-way ANOVA, n = 7 per group. B Protein expression of neurogranin (Ng), calmodulin (CaM), calcineurin (CaN), pan-CAMKII, and phospho-CAMKII (p-CaMKII) in mock and Ng-siRNA-treated hCMEC/D3 cells. There were no significant changes in the expression of CaM, CaN, CaMKII, and p-CaMKII following Ng-siRNA treatment in hCMEC/D3 cells C Protein expression change of claudin-1, claudin-5, occludin, zonula occludens-1 (ZO1), zonula occludens-2 (ZO2), and VE-Cadherin between mock and Ng-siRNA-treated hCMEC/D3 cells. Claudin-1, claudin-5, and zonula occludens-2 (ZO2) showed significantly reduced protein expression, while there were no changes in occludin, zonula occludens-1, and VE-cadherin expression ( n = 3). Data are mean ± SEM. * p < 0.05. Two-tailed unpaired Student’s t -test

Article Snippet: After transfection, the cells were digested with trypsin and plated on rat-tail collagen-coated cell culture polycarbonate transwell inserts (#3413, Corning) in the TEER24 gold electroarray plates (Applied biosciences) at a density of 10 5 per mL and grown endothelial cell media supplemented with FBS, ECGS and 10 U/ml penicillin, and 100 μg/mL streptomycin.

Techniques: Expressing, Permeability, Cell Culture, Control, Two Tailed Test