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. 2019 Jul 26;9:10888. doi: 10.1038/s41598-019-47267-5

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© The Author(s) 2019

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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BDL skin homogenate increased intracellular calcium through Mrgprx1. (a) Application of 2 µM BAM8-22 induced significant intracellular calcium increase, which implies that the transient expression of Mrgprx1 in HeLa cells was functional. (b) A schematic illustration of the experiment carried out for calcium imaging with skin homogenates. (c) Representative pseudo-coloured ratiometric images of sham or BDL skin homogenate treatment on HeLa cells expressing Mrgprx1 or pcDNA3.1 (“pcDNA”). (d) Time-course graphs of calcium imaging after treatment of skin homogenates collected from either sham or BDL mice on Mrgprx1- or pcDNA-transfected HeLa cells. The BDL skin homogenate significantly increased intracellular calcium level compared to that of sham. (e) Summary bar graphs showing the peak responses from either sham or BDL skin homogenate treatment onto Mrgprx1- or pcDNA-transfected HeLa cells. ***p < 0.001 (one-way ANOVA, Dunnett’s multiple comparison test). Results are presented as means ± SEM.