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. 2017 May 18;7:2090. doi: 10.1038/s41598-017-02242-w

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© The Author(s) 2017

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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Prediction of transcription factor binding sites and mutation analysis of the mSTING minimal promoter. (A) A schematic representation of the putative binding sites for DNA-binding proteins in the proximal promoter of the mSTING gene. The 129 nt DNA sequence (from −77 to +52) was analyzed with TFSEARCH softwares. The putative transcription factor binding sites were outlined with black box, and the names of the transcription were annotated. The transcription start site (TSS) was marked by black arrow. (B) Conserved base sequence of GATA1, IK2, Sp1/Sp3 or STAT binding site. The base size presented the binding affinity coefficient of transcription factor and promoter. (C) Mutation analysis of the mSTING minimal promoter. The left: Binding sites for GATA1, IK2, Sp1/Sp3 and STAT were indicated with open different shapes. Mutations were shown in bold above the histogram. The right: The relative luciferase activities derived from mutational pSTING-254. The site-special mutagenized plasmids were cotransfected with pRL-TK into NIH3T3 cells and lucifarase assays were performed. The level of firefly luciferase activities was normalized to the Renilla luciferase activity. Each bar represented the mean ± SD of three independent experiments. (n = 3, *p < 0.05 vs. pSTING-254).