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. 2021 Jan 19;12:439. doi: 10.1038/s41467-020-20714-y

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© The Author(s) 2021

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. 3 — a–f In situ hybridization against Fgf8 mRNA in e10.5 embryos from control (a, e), DEL64 (b), DEL79 (c), DEL80 (d) and DEL79-80 (f) mutants (n > 3 for all experimental groups). Note the complete lack of Fgf8 expression in the MHB of DEL64 (b) embryos as compared to controls (a). Brain tissue is markedly reduced already at this stage in DEL64 embryos (b). g, h In situ hybridization of control and DEL64 enhancer deletion embryos at e8.25 with riboprobe against Fgf8. No expression is detected in the DEL64 embryos (n = 4). i Relative expression of Fgf8 mRNA levels in dissected MHB region from WT (n = 11), DEL79 (n = 10), DEL80 (n = 12), and DEL79-80 (n = 3) e10.5 embryos as compared to Fgf8^null/+ (n = 11, 11, 10, 5) control littermates. Individual data points, mean ± SEM are indicated. *p < 0.05, n.s. = not significant (two-tailed Student’s t-test; exact p-values are indicated in the figure panel). Scale bar is 1 mm in (a–f) and 100 μm in (g, h).