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. 2021 Mar 29;11:7117. doi: 10.1038/s41598-021-86514-6

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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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Urolithin A diminished PI3K/AKT/mTOR activation in LPS-stimulated murine BMDMs. Murine BMDMs were stimulated by 1 µg/ml of LPS in the presence or absence of UA (25 µM or 50 µM) and harvested at indicated time intervals followed by western blot analysis. Phosphorylation of AKT (a, b) and mTOR (a, c) were monitored by immunoblot using pAKT (Ser473) and phospho-mTOR (Ser2448) monoclonal antibodies. Subsequently, blots were stripped and re-incubated with antibody against total AKT and mTOR. GAPDH served as a loading control. The unstimulated and untreated BMDMs were used as control. BMDMs treated with DMSO were used as negative control. Representative images and arithmetic means ± SEM from five independent experiments are depicted. Two way ANOVA was used and *(p < 0.05), **(P 0.01), ***(p < 0.001), and ****(p < 0.0001) indicate statistically significant differences compared to respective control. Full length blots are found in the Supplementary File 9. Abbreviations: DMSO, dimethyl sulfoxide; LPS, lipopolysaccharides; UA, urolithin A; GAPDH, glyceraldehyde-3- phosphate dehydrogenase.