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. 2020 Dec 17;41(3):1127–1145. doi: 10.1161/ATVBAHA.120.315031

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© 2020 The Authors.

Arteriosclerosis, Thrombosis, and Vascular Biology is published on behalf of the American Heart Association, Inc., by Wolters Kluwer Health, Inc. This is an open access article under the terms of the Creative Commons Attribution Non-Commercial-NoDerivs License, which permits use, distribution, and reproduction in any medium, provided that the original work is properly cited, the use is noncommercial, and no modifications or adaptations are made.

This article is made available via the PMC Open Access Subset for unrestricted re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the COVID-19 pandemic or until permissions are revoked in writing. Upon expiration of these permissions, PMC is granted a perpetual license to make this article available via PMC and Europe PMC, consistent with existing copyright protections.

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Figure 3. — Neural progenitor cell (NPC)-extracellular vesicles (EVs) reverse upregulation of ABCB1 (ATP-binding cassette subfamily B member 1 transporter) and activation of NF-κB (nuclear factor-κB) in endothelial cell (ECs) under both oxygen glucose deprivation (OGD) and lipopolysaccharide (LPS) conditions. A and B, Quantitative analysis of ZO-1 (zonula occludens 1) and ABCB1 expression in normoxia control, OGD and OGD treated with EVs using Western blot analysis normalized with the housekeeping protein α-tubulin (n=4 per group). C and D, Quantitative analysis of p65 and IκBα (inhibitor of nuclear factor-κB) expression associated with the NF-κB pathway in the same three groups using Western blot analysis normalized with the housekeeping protein α-tubulin (n=4 per group). E, Cell viability was analyzed in ECs exposed to 16 h of OGD followed by 24 h of reoxygenation using the MTT (methylthiazolyldiphenyl-tetrazolium bromide) assay (n=5 per group). Cells incubated under normoxic conditions were defined as 100% cell survival. F, The LIVE/DEAD assay uses the same conditions as mentioned for the MTT assay. The photos display representative immunofluorescence stainings of calcein AM (acetoxymethyl; LIVE cells, green) and ethidium homodimer-1 (DEAD cells, red). Scale bars: 200 µm (n=5 per group). G and H, Quantitative analysis of p65 and ABCB1 expression under control conditions and in ECs exposed to LPS 200, 500, or 1000 ng/mL using Western blot analysis normalized with the housekeeping protein β-actin or α-tubulin (n=4 per group). I and J, Quantitative analysis of p65 and ABCB1 expression in normoxia control, LPS (1000 ng/mL) and LPS-treated ECs in the presence of EVs using Western blot analysis normalized with the housekeeping protein β-actin or α-tubulin (n=4 per group). Data are expressed as mean±SD. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001.