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. 2022 Mar 9;12:4136. doi: 10.1038/s41598-022-07916-8

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© The Author(s) 2022

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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Effects of DNase treatment on NETs induction associated with muscle hyperalgesia. The MNT values of hindlimbs (a n = 8 in each group) and the uric acid level (b n = 6 in each group) in the TSM tissues in sham-stimulated control and stimulated mice injected intravenously with 10 mg/kg/day of DNase I or a vehicle. (c) Representative IHC images (200 ×) of the stimulated TSM tissue with or without DNase I administration on day 7, indicating citH3 (green), Gr-1 (red), and DAPI (blue). Scale bar = 50 μm. Western blotting analysis of citH3 (d) and Ly6G amounts in TSM tissues on day 7 (n = 6 in each group). The relative density is normalized based on the loading control (total H3 and GAPDH). (e) ELISA analysis was performed to evaluate protein levels of IL-1β in TSM tissues on day 7 (n = 12 mice without DNase treatment, n = 6 mice with DNase treatment). All data are shown as Tukey’s boxplots with individual data points and analyzed using unpaired t-test for comparing the mean of two different samples or two-way ANOVA followed by post-hoc Tukey’s test for a comparison of more than two groups. Statistical significance is indicated with * (p < 0.05), ** (p < 0.01) and *** (p < 0.001), **** (p < 0.0001).