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. 2018 Jul 30;8:11391. doi: 10.1038/s41598-018-29740-9

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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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Xbra and Smad-1 synergistically regulate ventx1.1 transcription. Different ventx1.1 promoter constructs were co-injected either with or without Smad-1 and with Xbra, either in combination or separately, at the 1-cell stage and grown until stage 11 to measure the relative promoter activity. (a,b) ventx1.1 (−103 and −180) injected with Xbra and Smad-1, in combination or separately. (c) XbRE-mutated ventx1.1 (−103) mXbRE and ventx1.1 (−103) injected with Xbra and Smad-1, in combination or separately, in separate groups. (d) XbRE and BRE were mutated by site-directed mutagenesis in different ventx1.1 (−180) promoter constructs. (e) XbRE-mutated ventx1.1 (−180) mXbRE injected either with or without Xbra and Smad-1, in combination or separately. (f) BRE-mutated ventx1.1 (−180) mBRE injected either with or without Xbra and Smad-1, in combination or separately. (g) The doubly mutated ventx1.1 (−180) m(BRE + XbRE) injected either with or without Xbra and Smad-1, in combination or separately. (h,i) Flag-Smad-1, Flag-Smad-1 (3SD), and Flag-Smad-1 (3SA) constructs were co-injected with Myc-Xbra and immunoprecipitation was performed with an anti-Flag antibody. (j) ventx1.1 (−180) injected either with or without Myc-Xbra, Flag-Smad-1, and Flag-Smad-1 (3SA) constructs in different groups, in combination or separately. All relative promoter activity data are shown as mean ± SE.