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. 2020 May 2;7:72. doi: 10.1038/s41438-020-0294-4

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© The Author(s) 2020

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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Fig. 8 — a The 1520-bp sequence of the MdTMT1 promoter. b GUS histochemical staining of a callus harboring a plasmid containing the MdTMT1 promoter and GUS reporter gene (TMT1 pro: GUS) and a callus overexpressing MdMYB6 harboring the TMT1 pro:GUS vector (TMT1 pro: GUS + MYB6). c Analysis of GUS activity. d Yeast one-hybrid analysis of the interaction between the MdYB6 protein and the MdTMT1 promoter. e Predicted elements bound by MdMYB6 in the MdTMT1 promoter. f ChIP-PCR analyses of the interaction between the MdTMT1 promoter fragments and the MdMYB6 protein. MdMYB6-DNA complexes were then coimmunoprecipitated from the calli overexpressing MdMYB6-GFP via a GFP antibody. g EMSA analysis of the interaction between the MdTMT1 promoter and MdMYB6 protein. Lane 1 included labeled DNA probes with no added protein, while lane 2 included labeled DNA probes with no competitor or protein. Unlabeled DNA probes were added in increasing amounts (25×, 50×, and 100×) in lanes 3, 4, and 5. Unlabeled mutant DNA probes (lanes 7, 8, and 9) were added as competitors. Lane 6 included labeled mutant DNA probes and protein but without competitor.