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. 2021 Jun 14;11:12465. doi: 10.1038/s41598-021-91937-2

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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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Effects of carbachol on heart rate in db/db mice in vivo. (A) Representative surface ECGs in baseline conditions and after application of CCh (0.1 mg/kg) in anesthetized wildtype and db/db mice. (B) Summary of heart rate at baseline and after CCh in wildtype (n = 9) and db/db (n = 13) mice. *P < 0.05 vs baseline; ⁺P < 0.05 vs. wildtype by two-way ANOVA with Holm–Sidak posthoc test. (C) Change in heart rate after CCh application in wildtype and db/db mice. *P < 0.05 vs. wildtype by Student’s t-test. (D) Time course of changes in heart rate after application of CCh and atropine (10 mg/kg) in wildtype (n = 5) and db/db (n = 6) mice. (E) Magnified region of data from panel D illustrating the changes in heart rate upon application of atropine in wildtype and db/db mice. For panels (D) and (E) *P < 0.05 vs. wildtype by two-way repeated measures ANOVA with Holm–Sidak posthoc test. (F) Recovery time for heart rate after application of atropine. *P < 0.05 vs. wildtype by Student’s t-test.