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Fig. 2 | Fluids and Barriers of the CNS

Fig. 2

From: Activation of NMDA receptors in brain endothelial cells increases transcellular permeability

Fig. 2

Activation of NMDA receptor induces translocation of molecules that are involved in transcytosis in bran endothelial cells. A Primary human brain endothelial cells were treated with NMDA (25 μM) for different time points and cells were fixed and stained with Clathrin heavy chain (Red). 3D reconstructed images were placed on the right side for better appreciation of clathrin heavy chain localization at the cellular edge. Magnified images from left panel are presented on the right panel in both. Scale bar indicates 75 μm and 25 μm for left and right panel, respectively. B Percentage of cells that is positive for the membrane localization of Clathrin heavy chains were quantified and depicted as a graph (n = 6, two-tailed unpaired student t-test, ** indicates where p < 0.01). C Activation of NMDA receptor induces redistribution of Caveolin 1 (Green). Scale bar indicates 75 μm and 25 μm. (D) AF594-transferrin permeability assay on the primary brain endothelial cells cultured on the porous membrane. Cells were pretreated with 10 μg/ml of AF594-transferrin with or without NMDA (25 μM) or MK801 (1 μM) and the transmigrated AF594-transferrin was measured using Fluorescence intensity of the AF594 conjugated transferrin that crossed the membrane were measured with excitation at 590 nm and emission at 617 nm with a Flex-Station 3 and its absolute concentration was calculated using standard curve. Values of transferrin concentration of treated groups were divided by those of control groups and percentage differences were presented as a graph (n = 4, two-way ANOVA, *, ** indicates where p < 0.05 and p < 0.01, respectively)

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