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. 2019 Mar 25;9:5043. doi: 10.1038/s41598-019-41639-7

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© The Author(s) 2019

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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Embryonic Hox status of periosteal stem/progenitor cells is preserved into adulthood. (A) Transcriptional map depicting normalized FPKM expression values for genes within the HoxA cluster. Note the near absence of Hox expression in the frontal and parietal bone, while proximal Hox genes are represented in the hyoid sample, and distal Hox genes are expressed in the tibia, similar to their embryonic pattern. Expression values from isolated periostea were averaged for each skeletal element (n = 3). (B) qPCR validation (mean +/− standard error) of three relevant Hox genes (Hoxa2, Hoxa11 and Hoxa13) identified as differentially expressed by RNA sequencing (n = 3). (C) In situ hybridization of hyoid and tibial periosteum with Hoxa2, Hoxa11 and Hoxa13 RNA antisense probes confirming spatial expression of the respective Hox genes within the periosteum (arrowheads)(n = 3). Abbreviations: c, cortical bone; p, periosteum.