Fig. 2

C9-ALS BMEC-like cells form a dysfunctional barrier A) Blood-Brain Barrier phenotypic markers. Ve-Cadherin (CDH5), Claudin-5 (CLDN5), Junctional Adhesion Molecule 2 (JAM2), Occludin (OCLN), Transforming growth factor beta 1 (TGFB1), Zonula Occludens 1 (TJP1) and, Von Willebrand factor (VWF), P-glycoprotein (ABCB1), EAAT3 (SLC1A1), EAAT2 (SLC1A2), EAAT1 (SLC1A3), Insulin receptor (INSR) and, receptor for advanced glycation end products (RAGE) transcriptional expression of 2 healthy donors and 3 C9-ALS donors hi-PSCs derived BMEC-like cells. qRT-PCR data are plotted as mean ± s.d. Statistical significance was determined using Student’s unpaired t-test (****p < 0.0001). N = 3 per group. B) Brightfield images are shown as follows: HUVECs, CTR and C9-ALS BMEC-like cells. The scale bar equals 50 µM. C) BMEC passive barrier as shown by TEER following subculture for GM23338 male healthy donor hi-PSCs derived BMEC-like cells (CTR BMECs), CS52iALS-C9nxx and CS29iALS-C9nxx male C9-ALS donors (ALS BMECs). Error bars represent the standard deviation of triplicate Transwell™ filters. Statistical significance was determined using Student’s unpaired t-test (****p < 0.0001). D) BMEC-like cells were incubated with or without Cyclosporin-A, a P-glycoprotein inhibitor and, next with Rhodamine-123, a P-glycoprotein fluorescent substrate. Accumulation is normalised to the no-inhibitor samples. Error bars represent the standard deviation of triplicate wells. Statistical significance was determined using One-Way ANOVA (****p < 0.0001). N = 3