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. 2021 Sep 15;11:18345. doi: 10.1038/s41598-021-97869-1

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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DCFDA staining of CSE-treated, SCAL1-deficient and SCAL1-overexpressing A549 cells to assess production of reactive oxygen species (Olympus IX51, ×100, FITC: 500 ms). (a,b) The fluorescein-based dye DCFDA exhibits green fluorescence upon oxidation by ROS. The bright green signal observed in A549 cells signifies ROS production. DCFDA intensities normalized against cell count showed increased mean ROS production upon SCAL1 knockdown. (c,d) DCFDA fluorescence signals were lower in the SCAL1 overexpression setup, suggesting that increased SCAL1 levels decrease ROS production. Data presented are representative of three independent trials performed in triplicates and are expressed as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001.