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. 2018 Jun 14;9:447. doi: 10.3389/fneur.2018.00447

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Copyright © 2018 Avezov, Aizenbud and Lavie.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Production of Reactive oxygen species (ROS) by EC-CFUs under normoxia, intermittent and sustained hypoxia with and without diphenyl iodide (DPI). Production of ROS per colony was measured by the nitro blue tetrazolium (NBT) test with and without the NADPH oxidase inhibitor DPI (5 μM) (n = 3). The test depends on the direct reduction of NBT into an insoluble blue formazan compound by NADPH oxidase. Cytoplasmic clumps of formazan deposits are considered positive for ROS. The blue score positively correlates with cellular ROS production. (A) EC-CFU formazan optic density on the 7th day in culture with and without DPI under normoxia (Norm), intermittent hypoxia (IH) and sustained hypoxia (SH) (p < 0.01 for: Norm vs. Norm+DPI, SH vs. SH+DPI; p < 0.1 for IH vs. IH+DPI). (B) Representative photomicrographs of EC-CFUs microscopic fields (X10) of NBT test for Norm, IH and SH with and without 5 μM DPI.