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. 2020 Jul 14;18:87. doi: 10.1186/s12915-020-00813-4

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — Cell fate change and ectopic renewal of Sox2(+) cells in the dental epithelium of K14^Cre/+;Fam20B^fl/fl mice. A–M GAG deficiency in the dental epithelium led to ectopic over-proliferation and less apoptosis in the native incisors. EdU incorporation analysis showed ectopic over-proliferation (white arrows in F–J) in the GAG-deficient dental epithelium (e, plotted by dashed lines) and dental mesenchyme (m) at the mesial/lingual side of native incisors in the presumptive location of supernumerary teeth formation starting at E13.5 compared to the normal control mice (2-sample t tests, P < 0.05). TUNEL assay revealed reduced apoptosis (yellow arrow) in GAG-deficient dental epithelium at E12.5 (2-sample t tests, P < 0.01). N–Q Three-dimensional reconstruction of lower incisor confocal images after tissue clearing of mandibles. td^Tomato indicates the dental epithelium. GFP indicates the Sox2(+) cells. On the sagittal sections of E15.5 incisors (N, O), an ectopic thickening of dental epithelium was identified at the lingual side of the native enamel organ in K14^Cre/+;Fam20B^fl/fl mice (arrow) compared to the normal control. The slight bulge (white arrow) at the lingual side of the enamel organ in the Fam20B-mutant mice indicates the initiation of the supernumerary tooth outgrowth. At E16.5 (P, Q), Sox2(+) cells showed ectopic renewal in the Fam20B-deficient native enamel organ and actively contributed to the development of successive enamel organ (arrow). In contrast, the normal controls had lost Sox2 expression at this stage. The dashed lines in (P) and (Q) outlined the remaining parts of the enamel organs that were not able to be displayed due to the working distance of the confocal. Scale bars: 25 μm in (A–J); 50 μm in (N–Q)