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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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Elimination of the induced expression of ZIPs results in a reduced expression of cell-surface ZNT1. (A) Schematic depicting the assay time course. MDCK cells stably expressing Zip4, Zip5, ZIP4, or ZIP5 were cultured with or without 0.1 μg/mL Dox for 24 h, and then the cells were cultured in normal medium (N) for an additional 24–72 h after washing thrice with PBS. (B,C) ZNT1 expression upregulated by Dox-induced expression of Zip4 or Zip5 (B) or ZIP4 or ZIP5 (C) decreased after Dox removal. However, a marked reduction of MT expression did not accompany the reduction of ZNT1 expression. Actin was used as the loading control. Each experiment was performed at least thrice, and representative results from independent experiments are shown.