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. 2017 Aug 2;7:7129. doi: 10.1038/s41598-017-06310-z

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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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Craniofacial skeletal abnormalities in Irf6 null pups and quantitative expression of selected genes. (A–H) Bone (red) and cartilage (blue) of head and mandible of P0 pups. Relative to wild type littermates (A,C,E,G), mutant pups (B,D,F,H) have less cartilage (B,F), and the mandible is missing a distal bone where the incisor is normally formed (E versus F). The mandibular processes are shorter (F). The coronal sutures of the skull of Irf6 null pups are fused (arrows) (D,H) compared to wild type littermate (C,G). Scale bars: 1000 μm. Expression of Irf6, Tw1 and Runx2 is reduced in Irf6 null embryos at E14.5 compared to wild type littermate (I–K). Similarly, the expression of Irf6 and Runx2 genes is reduced to a lesser extent in compound alleles for Irf6 null and p53 heterozygous embryos at E14.5, except for Tw1 where the expression is slightly increased (J). The relative gene expression is the average of two independent experiments with four technical replicates for each genotype. The asterisks represent the level of statistical significance of the average of gene expression compared to wild type embryos. The data are the average of two independent experiments. Similarly, the expression of Irf6 and Grhl3 is significantly reduced in Irf6 null embryos at both E10.5 and E12.5 (L,M).