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. 2019 Mar 5;9:3585. doi: 10.1038/s41598-019-40329-8

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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GCs are a source of H₂O₂ and other ROS produced by NOX4. Basal production of ROS (identified by the indicator dye H₂DCFDA, n = 8) and specifically of H₂O₂ (detected by Amplex^® Red, n = 4) was measured on culture day 2 to 4 for 2 h (A,B). For clarity, results are presented as mean only (six technical repetitions) and as fold-change relative to the intensity at t = 0. The NOX4 blockerGKT137831 (20 µM) reduced the production of ROS/H₂O₂. One representative measurement for both is shown in (C,D). Blocking NOX4 significantly reduced production of ROS (E; by 55%; n = 6) and H₂O₂ (F; by 36%; n = 4) after 2 h (mean ± SEM). One-sample t-tests (two-tailed; *p < 0.05; ***p < 0.001).