Astrocyte conditioned media (CM–ASTs) induce brain endothelium properties on HUVEC.

<p>HUVEC were culture with CM-ASTs (HUVECd) or complete medium DMEM (HUVEC) for 24 h. Thereafter, alkaline phosphatase (AlkP) activity (<b>A</b>) or transendothelial monolayer passage of horse radish peroxidase (HRP) (<b>B</b>) were spectrophotometrically assessed. Occludin (<b>C</b>) and ZO-1 (<b>D</b>) expression was FACS analyzed and cellular expression of ZO-1 and distribution of F-Actin were evaluated by microscopy (<b>E</b>) as described in M&M. <b>A</b>: Enzymatic units (EU) of AlkP specific activity (Mean ± SEM). * p<0.05 (CM-ASTs vs. DMEM, n = 4). <b>B</b>: % Relative permeability (Mean ± SEM, n = 3) of endothelial monolayer to HRP. <b>C and D</b>: Occludin and ZO-1 relative fluorescence (RF, median fluorescence intensity (MFI)/isotype control MFI) (Mean ± SEM).*p<0.05 (CM-ASTs vs. DMEM, n = 3). <b>E</b>: Photomicrography showing ZO-1 expression pattern of HUVEC (upper left panel) and HUVECd (upper right panel) of one representative experiment (n = 3). Photomicrography showing maximum intensity Z-projection of F-Actin distribution on HUVEC (lower Left panel) and HUVECd (lower right panel) of one representative experiment (n = 4).