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. 2020 Feb 6;10:1962. doi: 10.1038/s41598-020-58999-0

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© The Author(s) 2020

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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H₂ regulates sirtuin expression. Mice in Con, HRS, OVA or OVA + HRS group were sensitized and challenged, and treated with HRS as described above. Lung tissue levels of sirtuins (Sirt) 1, 2, 3, 4, 5, 6 and 7 mRNA expression were quantified by qRT-PCR and expressed as sirtuin/GAPDH (glyceraldehyde 3-phosphate dehydrogenase) ratio, or lungs tissue levels of sirtuins 1, 3 and 6 proteins were quantified by Western blots. Means ± SEM of 5 mice per group. *p < 0.05, compared to control group. ^#p < 0.05, compared to OVA group (unpaired t test or one-way ANOVA). (A) Sirtuin mRNA expression in control lungs. (B) Sirtuin mRNA expression in OVA-sensitized and challenged lungs. (C–E) Sirtuins 1, 3 and 6 protein expression in OVA-sensitized and challenged lungs. The Western blot bands (upper panel) were quantified using densitometry and expressed as fold change over control group (bar graphs). Cropped blots are displayed. The original blots are shown in Supplementary Fig. 2A–C.