Fig. 2: Blood–brain barrier formation induced by brain-derived ECs.
From: Brain-derived endothelial cells are neuroprotective in a chronic cerebral hypoperfusion mouse model
Vascular phenotypes in brains transplanted with ECs from different organs were analyzed on day 21 as described in Fig. 1a. a Representative images show staining with anti-CD31 mAb (Alexa Fluor 647) and anti–claudin-5 pAb (Alexa Fluor 546) in regenerated vasculature transplanted with GFP-positive ECs derived from organs as indicated. The positive rate refers to the proportion of claudin-5–positive ECs among GFP-positive ECs expressed as a percentage. Scale bar, 30 μm. n = 5 per group. Data are presented as mean ± SEM. b, c Representative images stained with anti-CD31 mAb (Alexa Fluor 647), anti–NG2 pAb (Alexa Flour 546) (b), anti-CD31 mAb (Alexa Flour 647), and anti-GFAP pAb (Alexa Flour 546) (c) in regenerated vasculature areas transplanted with GFP-positive ECs derived from organs as indicated. The positive rate refers to the proportion of NG2-positive pericytes covering ECs (b) or astrocyte end-feet (GFAP-positive) positive ECs (c) among GFP-positive ECs expressed as a percentage. Scale bar, 30 μm. n = 5 per group. Data are presented as mean ± SEM. d Fluorescence images of vasculature transplanted with GFP-positive ECs derived from various organs as indicated. Blood flow was overlapped by labeling with AngioSPARK 680. The positive rate refers to the proportion of GFP-positive vessels with blood flow among GFP-positive ECs expressed as a percentage. Scale bar, 300 μm. n = 3 per group. Data are presented as mean ± SEM. *, P < 0.05; ***, P < 0.001; ****, P < 0.0001. EC endothelial cell, GFAP glial fibrillary acidic protein, GFP green fluorescent protein, mAb monoclonal antibody, NG2 neural/glial antigen-2, pAb polyclonal antibodies.
