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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⁺ successfully increases the surface AQP-5 level on the plasma membrane via ZnR/GPR39 activation. HSG cells cotransfected with shRNA and AQP-5-Myc were treated with vehicle, 300 µM carbachol, or 100 µM ZnCl₂ for 15 secs (0.25 min) and membrane translocation of AQP-5 was analyzed. (a,b) Confocal images of shControl-transfected HSG cells (a) and shGPR39-transfected HSG cells (b) stained with total AQP-5 (blue) and surface-AQP-5 (red) and expressed with GFP (green). Merged image of total and surface-AQP-5 was depicted (purple). Scale bar, 5 μm. (c,d) Quantification of surface-to-total AQP-5 ratio in shControl- (c) and shGPR39-transfected HSG cells (d). Values are presented as percentages of vehicle. The experiments were performed three times independently, and the results were reproducible. Numbers of counted cells were indicated inside bars. ***P < 0.001, compared with vehicle control. n.s., not significant.