Fig. 9

High circulatory Lcn2 causes blood brain barrier dysfunction. a, b Immunoreactivity of Claudin 5 (green) in a and Zonula Occludens (ZO1) (Red) in b as shown by immunofluorescence microscopy in mouse primary brain endothelial cells exposed with or without siRNA-24p3R and treated with VEHICLE (DMSO), Mouse recombinant Lcn2 (100 ng/ml), 20 μl of serum from CHOW, and MCD fed mouse. Nucleus was stained with DAPI. Images were taken at × 20 magnification. c, e Morphometry analysis of Claudin 5 (c) and ZO1 (e) immunoreactivity in brain endothelial cells without exposure of 24p3R siRNA. Immunoreactivity was displayed as bar graph representing mean ± SD of ROI % of 3 different areas (n = 3). d, f Morphometric analysis of Claudin 5 immunoreactivity in d and ZO1 in f in primary brain endothelial cells exposed with or without 24p3RsiRNA. Data was calculated as mean ± SD of %ROI of 3 different fields and displayed as bar graph. g, h IL-6 and Il -1β levels in pg/ml in supernatants from brain endothelial cells were displayed by bar graph. Data was calculated as expressed as mean ± SD, and significance was calculated by paired t test between the groups, *p < 0.05, **p < 0.01, ***p < 0.001. All statistical analysis was performed by unpaired and paired t test between the means. Significance was tested by paired test between the groups. *p < 0.05, **p < 0.01, ***p < 0.001, followed by Bonferroni post-hoc corrections