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Fig. 4 | Journal of Neuroinflammation

Fig. 4

From: Gene expression comparison reveals distinct basal expression of HOX members and differential TNF-induced response between brain- and spinal cord-derived microvascular endothelial cells

Fig. 4

TNF-α effects on MEC monolayers integrity (24 h incubation). a Representative photomicrographs of BMEC monolayers fixed, permeabilized, and stained with an antibody directed against occludin (scale bars, 10 μm). Nuclei are stained with Hoechst. Arrows indicate pericellular localization of TJ protein occludin, while arrowheads point to cytoplasmic/vesicular of occludin. Note the decreased pericellular and increased cytoplasmic/vesicular distribution of occludin upon TNF-α treatment. b Western blot analysis (left panel) and quantification (right panel) of TJ proteins ZO-1, occludin, and claudin-5 levels were performed in whole tissue lysates from BMEC monolayers. Quantification of the optical densities of each band was assessed using the NIH ImageJ software. Values deduced from non-treated cultures (Ctrl) were considered as 100 %. c Barrier integrity was assessed by quantification of the Lucifer Yellow (LY) endothelial permeability, Pe(LY). The loss of barrier integrity was validated for Pe(LY) above 0.6.10−3 cm/min. All data are representative of at least three experiments with similar results and are presented as means ± standard deviation (**p ≤ 0.01; ***p ≤ 0.001)

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