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. 2023 Jun 7;475(7):807–821. doi: 10.1007/s00424-023-02825-9

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© The Author(s) 2023

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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Fig. 5 — Acrolein effects on extracellular ROS formation on RAW and EA.hy cells. RAW cells (A) and EA.hy cells (B) were incubated with dihydroethidium (DHE) and the extracellular oxidation products 2-hydroxyethidium (2-HE, superoxide specific) and ethidium (E + , unspecific) in the cell culture supernatant were separated by HPLC. Specifically phagocytic NADPH oxidase produces high amounts of extracellular superoxide in immune cells. Concentrations of 2-HE and E + are shown with the representative chromatograms. Each data point indicates an independent experiment. Data are presented as absolute concentration (mean ± range). Conventional two-tailed t-test was used for statistical analysis. C Luciferase reporter assay was used to evaluate the activation of Nrf2 in acrolein-treated DLD1-HO1-prom cells. D The scheme summarizes the principles of the assay. Chemiluminescence after treatment with different concentrations of acrolein is shown as a percentage of control (mean ± range, n = 8 independent experiments). Conventional 1-way ANOVA was used for statistical analysis. Significance is indicated as *, **, *** and **** when p < 0.05, p < 0.01, p < 0.001 and p < 0.0001 respectively