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. 2019 Nov 27;9:17648. doi: 10.1038/s41598-019-54173-3

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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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Zn²⁺ evokes [Ca²⁺]_i increase via ZnR/GPR39 in human salivary gland cells. (a,b) left, Fura-2-loaded HSG cells (a) and A253 cells (b) were challenged with ZnCl₂ at various concentrations (1, 10, 100 µM) and then monitored for changes in the fluorescence ratio of F340/F380 to represent [Ca²⁺]_i level. right, Concentration–response relationships were also depicted by measuring the peak height of changes in [Ca²⁺]_i. A sigmoidal fitting curve is also depicted (gray). (c,d) Fura-2-loaded HSG cells transfected with siControl and siGPR39 were challenged with 200 µM ZnCl₂ (c) or 300 µM carbachol (d) and then monitored for changes in the fluorescence ratio. Quantification of the change of [Ca²⁺]_i mediated by ZnCl₂ and carbachol was normalized to control groups. The experiments were performed three times independently, and the results were reproducible. ***P < 0.001, compared with control.