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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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Hormonal stimulation significantly increases levels of DUOX, but does not elevate basal ROS production in GCs. Results of qRT-PCR experiments showed that mRNA expression levels of NOX5/DUOX1/2 were elevated after hormonal stimulation, but not all reached statistical significance (A,B). Results (mean ± SEM) shown were normalized to the geometric mean of the reference genes RPL19, UBC, PPIA and GAPDH. (C) Measurements of ROS production using H₂DCFDA did not show any difference after 2 h measurement when treated with the hormones. Numbers of repeated experiments are shown inside the columns. Mean ± SEM; one-sample t-tests (two-tailed; *p < 0.05)