Fig. 4

VCE-004.8 prevents BBB disruption in vitro. A VCE-004.8 treatment protected against endothelial disruption promoted by PBMCs supernatant in the bEnd5 cell line. Data represent the mean ± SD (n = 3) and significance determined by one-way ANOVA followed by Tukey’s test. p = 0.0007; ***p < 0.001 for PBMCs supernatant-treated cells vs control; p = 0.0007; ###p < 0.001 for VCE-004.8-treated cells vs PBMCs supernatant-treated cells. B, C Representative fluorescence images of bEnd5 cells cultured in Transwell filters from the TEER assay stained for CLDN5 and ZO-1 tight junction proteins. A representative image of three independent experiments is shown (n = 3). Scale bars equivalent to 25 μm. Representative images of immunostaining of VCAM1 (D) and ZO-1 (E) expression in cultured bEnd5 cells stimulated during 24 h with proinflammatory cytokines. Cells were counterstained with DAPI to identify nuclei. A representative image of three independent experiments is shown. Scale bars equivalent to 25 μm