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. 2017 Sep 5;8:436. doi: 10.1038/s41467-017-00100-x

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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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Fig. 5 — In vivo validation of putative driver mutations in DHS chr6:28948439-28951450. a Shown are the locations of the four mutations in GATA and ETS transcription factor binding sites that were tested for function. b Barplots showing downregulation in anterior neural plate (a6.5 lineage, chr6:28949254 A > C), no change in endoderm (chr6:28950040 C > T), downregulation in endoderm (chr6:28950050 G > A) and overexpression in head epidermis (chr6:28950885 A > G). p-values were calculated using Fisher’s exact test. (c–g) Images showing tailbud stage C. intestinalis embryos (8 h post fertilization at 21 degrees Celsius) electroporated with (c, f) indicated reference enhancer > GFP or (d, e, g) indicated mutated enhancer > GFP. d Decreased expression in the anterior neural plate is observed when the enhancer is mutated (chr6:28949254 A > C); e Enhancer activity is decreased in anterior neural plate and secondary notochord in presence of mutation chr6:28950050 G > A. c is the wild-type enhancer for both d and e. g Increased expression in the epidermis is observed when the enhancer is mutated (chr6:28950885A>G)