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. 2018 Jan 12;8:593. doi: 10.1038/s41598-017-18911-9

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© The Author(s) 2018

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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Confirmation of loss-of-function of the VDR gene in the knocked-in cell lines. (a) Immunoblot analyses of isolated clones. αHsp90 and αβ-ACT were used as loading controls. (b) RT-qPCR analysis of CYP24A1 of the isolated cells with or without treatment with 1α,25-dihydroxyvitamin D3 (1,25D3). (c) Promoter-reporter assay using vitamin D response element (VDRE). When endogenous VDR was functional, VDRE would respond to 1,25D3 treatment. The exogenous VDR expression plasmid (VDR+) was transfected for the complementation assay.