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. 2020 Dec 3;10:21167. doi: 10.1038/s41598-020-78398-9

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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 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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Intravesical ATP treatment triggers activation of L6–S1 spinal neurons and induces an enhanced micturition reflex. (a) Representative cystometrograms after ATP instillation in wild-type mice. Scale bar, 10 min. (b) Changes in ICIs after ATP treatment (n = 5 mice per group); *p = 0.041 for 20 mM ATP, *p = 0.010 for 50 mM ATP, and **p = 0.0018 for 100 mM ATP (paired t-test with Holm correction). (c) Ultrasonographic findings of pre-voiding with intravesical ATP treatment. Scale bars, 5 mm. (d, e) Changes in largest CSA (d) and smallest CSA, post-voiding (e) (n = 5 mice per group); *p = 0.041 in saline versus 5 mM ATP, and ***p < 0.001 in saline versus 20 mM ATP and 50 mM ATP (Tukey’s test following one-way ANOVA). (f) Immunohistochemical analysis of c-Fos expression in L6–S1 spinal cord after ATP instillation. Scale bars, 100 μm. (g) The distribution of c-Fos-positive cells in the L6–S1 spinal cord induced by ATP instillation (50 mM) in wild-type mice (n = 5 mice per group); ***p < 0.001 in SPN and DCM (Student’s t-test with Holm correction). (h) The number of c-Fos-positive cells in L6–S1 spinal cord after ATP instillation (n = 5 mice per group: 10 sections per mouse were assessed). ***p < 0.001 in saline versus all ATP concentrations (Tukey’s test following one-way ANOVA). Error bars represent s.e.m., n.s., not significant.