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. 2018 Oct 16;12:359. doi: 10.3389/fncel.2018.00359

FIGURE 1.

FIGURE 1

PX dose-dependent increase in toxicity and Pe of the human BBB model. (A) Schematic drawing of the in vitro BBB co-culture system applied in this study: co–culture model of BLEC grown on matrigel-coated Transwell® permeable inserts with bovine brain pericytes grown at the abluminal side in a non-contact manner, used for functional Pe experiments. (B–D) Monolayers of BLECs were treated with PX for 24 h in a dose response experiment. Viability and cell death were examined by (B) MTT assay –a representative (out of three) experiment, (C) LDH release (n = 37-46 from five independent experiments), and (D) cytotoxgreen staining (a representative figure, out of three time course independent experiments is shown, n = 4–6 for each time point), the relative fraction of dead cells out of the whole population is presented as the ratio of green object confluence to phase confluence, ∗marks the first time in which the PX treatment is significantly different from control according to plot color, respectively. (E,F) Co-culture BBB in vitro model was treated with PX at the luminal side for 24 h in a dose response experiment. (E) To assess TJ functionality, Pe of sodium fluorescein (NaF) across the BBB in vitro model (from luminal to abluminal side) was assessed (n = 11–25 from four independent experiments) and cell death was examined by LDH release at the luminal (F) and abluminal (G) side (n = 11–25 from four independent experiments). Data presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01 and ∗∗∗p < 0.001 vs. control (Ordinary one way ANOVA with Dunnett’s multiple comparisons test).