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. 2024 Jun 5;14:12874. doi: 10.1038/s41598-024-62828-z

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© The Author(s) 2024

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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The effect of ESS on cell viability and anti-inflammatory responses in Raw 264.7 cells. The cell viability of RAW 264.7 cells was assessed by (a) treating with concentrations of ESS (2.5, 5 ppm) for 24 h and (b) pretreating with concentrations of ESS (2.5, 5 ppm) for 1 h, followed by treatment with LPS 500 ng/mL for 24 h. Based on the cell viability result, the anti-inflammatory effects of ESS on (c) No assay, (d) IL-6, and (e) TNF-α mRNA expression were determined. The RT-PCR analysis for RAW 264.7 cells was conducted in 12-well plates (4 × 10⁵ cells/well), with cells at 80% confluency pretreated with concentrations of ESS (2.5, 5 ppm) for 1 h, followed by treatment with LPS at 500 ng/mL for 6 h. The values presented are the mean ± SEM obtained from at least three independent experiments. Statistical significance is indicated as follows: ***p < 0.001, **p < 0.01, and *p < 0.05.