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. 2018 Jan 30;8:1838. doi: 10.1038/s41598-018-20216-4

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© The Author(s) 2018

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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Pharmacological properties of BSM contractility. BSM strips were stimulated by EFS at indicated frequencies in increasing order ((A)-left). Raw data showing effect of sequential addition of atropine and NF449 (pre-treatment 15 min) on EFS force ((A)-right panels). Selective P2X₁ antagonists NF449 ((B), n = 7), NF279 ((C), n = 4), or selective P2X₄ antagonist 5-BDBD ((D), n = 7), or P2X₄ potentiator, Ivermectin ((E), n = 8) were then added at indicated concentrations (with accumulative addition) for 15 min to measure the atropine-resistant force change in BSM contraction in response to EFS. Force changes were normalized to control and shown as percentages. These data indicate that P2X₁ and P2X₄ antagonists only partially inhibit BSM atropine resistant force at relatively high concentrations. Ivermectin, a P2X₄ potentiator, does not increase BSM atropine resistant force significantly. BSM atropine resistant force before drug treatment was used as control (100%), and drug effect (inhibition or potentiation) was normalized to control. Data were analysed using one-way ANOVA and then Bonferroni’s multiple comparison post-hoc tests. * indicates P < 0.05 when compared to control.