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. 2018 Feb 16;8:3155. doi: 10.1038/s41598-018-21194-3

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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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Promoter activity of mouse Tnmd in rat tenocytes. (a) Various lengths of DNA fragments of mouse Tnmd (175–1114 bp) in the forward or reverse orientation were cloned into the promoterless pGL4.10[luc2](pGL4.10 basic) Vector or pGL4.23[luc2/minP](pGL4.23) with a minimal promoter. The location of the first nucleotide of the upstream transcription start site is denoted as +1. The putative E-box sites and TATA box are shown by grey and black boxes, respectively. (b,c) Promoter activities of the mouse Tnmd gene in Tnmd-expressing tenocytes. Cells were co-transfected with a series of constructs shown in (a) and pGL4.74[hRluc/TK]. Firefly and Renilla luciferase activities were measured 24 h after transfection. The relative luciferase activity normalized to the Renilla luciferase activity is depicted as the fold-induction compared to each activity of the empty vector. All dual luciferase assays were performed in triplicate. The graphs show one representative experiment out of three. Each bar represents the average of three independent transfections (means ± SD.).