Fig. 2

HD180 iBMECs exhibit altered protein localization, gene expression, and BBB function. A Immunofluorescence of BMEC proteins (claudin-5, VE-cadherin, ZO1, occludin, ZO1, P-glycoprotein). Representative images of iBMECs are shown at 2 days after seeding. Inset shows mislocalization of ZO-1 at higher magnification for HD180 iBMECs. B Semi-quantitative analysis of iBMEC protein expression at day 2 following differentiation. The fluorescence signal was normalized to the nuclear DAPI signal and then plotted relative to HD-corrected. Data collected across n = 4–6 independent differentiations. C, D TEER time course and average over 10 days. Data collected across n = 16 (HD-corrected) and 26 (HD180) independent differentiations. E, F Lucifer yellow and 10 kDa dextran permeability (day 2 and day 10). Data collected across n = 6 (HD-corrected) and 8 (HD180) independent differentiations. G Rhodamine 123 efflux ratio (day 2). Data collected across n = 4–5 independent differentiations for HD-corrected and HD180 iBMECs. H Glucose permeability (day 2). Data collected across n = 4 independent differentiations for HD-corrected and HD180 iBMECs. All recordings in C–G represent averages across n = 2–6 technical replicates (individual Transwells) for each biological replicate