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. 2022 May 5;12:7334. doi: 10.1038/s41598-022-10925-2

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© The Author(s) 2022

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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ZNT1 and MT expression increases in a coordinated manner in response to Dox-induced expression of Zip4/Zip5 or ZIP4/ZIP5. (A) Upregulated expression of ZNT1 and MT accompanied Dox-induced expression of Zip4/Zip5 in MDCK cells. MDCK cells stably expressing Zip4 or Zip5 were cultured with the indicated concentrations of Dox for 24 h, and then ZNT1 and MT expression was examined. Tubulin was used as a loading control. (B,C) ZNT1 accumulated on the cell surface upon Dox-induced Zip4/Zip5 expression. MDCK cells cultured with or without 1.0 μg/mL Dox for 24 h were used in cell-surface biotinylation assays (B) or subject to immunofluorescence staining (C). In (B) tubulin and Na⁺/K⁺-ATPase were used as the loading control for input and biotinylation, respectively. (D–F) Same experiments as in (A–C) but performed using ZIP4 and ZIP5. Each experiment was performed at least thrice, and representative results from independent experiments are shown.