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. 2020 Feb 3;10:1656. doi: 10.1038/s41598-020-58222-0

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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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Immunohistochemical staining, species specific FISH probe staining and alizarin red staining of complex cell sheet. (A) MC3T3-E1 cell sheet showed positive staining for anti-mouse-OCN antibody, (B) PDL cell sheet did not show any positive staining for anti-mouse-OCN antibody, (C) Complex cell sheet showed positive expression for anti-mouse-OCN antibody in MC3T3-E1 cells space and negative staining in the PDL cells area, PDL cells and MC3T3-E1 cells area is demarcated using black dotted lines, (D) Rat species specific FISH probe stained complex cell sheet shows positively expressed rat PDL cells in red color, cells counterstained with DAPI are shown in blue, PDL cells and MC3T3-E1 cells area is demarcated using yellow dotted lines, (E) Alizarin red staining of complex cell sheet did not show any mineral deposition.