Fig. 2

Isolated endothelia are proliferative and re-establish blood-brain barrier phenotype. Endothelial and pure pericyte cultures were treated with EdU (48 h, 10 μM) then fixed and stained for the junctional proteins ZO-1, claudin-5, CD31, endothelial nuclear marker ERG, and proliferation marker EdU. RNA was extracted from endothelia, and a single pure pericyte culture, and gene expression analysed by qPCR. a Representative images of immunocytochemical staining of zona occludens-1 (ZO-1), CD31, claudin-5, and ERG in endothelia and pericytes. Scale bar = 500 um. b High-magnification representative images of co-staining of CD31 and claudin-5, and c CD31 and claudin-5. Scale bar = 25 μm. d Gene expression analysis of endothelial cultures (mean ± SEM, n = 3), normalised to gene expression of a single pure pericyte culture. e Representative images of EdU staining in endothelia and f quantification of basal proliferation over 48 h