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. 2020 Jan 2;8:1. doi: 10.1038/s41413-019-0078-3

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© The Author(s) 2020

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. 6 — Altered WNT/β-catenin signaling in calvaria from Dhcr7 and Insig1/2 mutant mice during craniofacial development. a Quantitative RT-PCR for Axin2 in calvaria from wild-type (WT; blue bars) control and Dhcr7 knockout (KO; red bars) mice at E14.5, E16.5, and P0 (newborn). n = 6 per group. *P < 0.05; ***p < 0.001. b Quantitative RT-PCR for Axin2 in calvaria from newborn WT (blue bar) and Insig1/2 conditional KO (cKO; yellow bar) mice. n = 6 per group. ***P < 0.001. c Quantitative RT-PCR for Axin2 after treatment with LiCl (left panel) or WNT3A (right panel) in WT (blue bars) and Dhcr7 KO (red bars) osteoblasts. n = 6 per group. ***P < 0.001. d Quantitative RT-PCR for Axin2 after treatment with LiCl (left panel) or WNT3A (right panel) in WT (blue bars) and Insig1/2 cKO (yellow bars) osteoblasts. n = 6 per group. ***P < 0.001. e β-galactosidase staining (blue) for sites of WNT/β-catenin signaling activation in the frontal bones of P0 Dhcr7^+/+;Topgal and Dhcr7^−/−;Topgal mice. Nuclei were stained with nuclear fast red. Scale bars, 100 µm. f Hematoxylin and Eosin staining of the sagittal sutures of newborn WT, WT;Axin2^L/+, Dhcr7 KO and Dhcr7 KO;Axin2^L/+ mice. Arrowheads indicate the osteogenic front. Accelerated bone formation of the sutures (frontal, coronal, and sagittal sutures) was normalized in newborn Dhcr7 KO;Axin2^L/+ mice (n = 6/6). Scale bars, 100 µm. g Von Kossa staining of the sagittal sutures of newborn WT;Axin2^L/+, Dhcr7 KO and Dhcr7 KO;Axin2^L/+ mice. Scale bar, 200 µm. h Quantitative RT-PCR of the indicated genes in newborn WT (blue bars), Dhcr7 KO (red bars), WT;Axin2^L/+ (gray bars) and Dhcr7KO;Axin2^L/+ (black bars) mice. n = 6 per group. ***P < 0.001.