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. 2021 Jan 5;8:572356. doi: 10.3389/fbioe.2020.572356

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Copyright © 2021 Anderson-Baron, Liang, Kunze, Mulet-Sierra, Osswald, Ansari, Seikaly and Adesida.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Alizarin Red S staining, COL X and CD31 immunofluorescence is reduced in pellets treated with PTHrP after 3 weeks of implantation in nude mice. After 3 weeks of implantation in nude mice, pellets were harvested, fixed in 10% formalin, sectioned, subjected to Alizarin Red S staining, and probed with fluorescent antibodies to COL X and CD31. Sections were imaged by brightfield/fluorescent microscopy. The cell source nasal chondrocytes (NC), bone marrow-derived mesenchymal stem cells (BM-MSC), or coculture (CC) and PTHrP treatment is indicated on top of the images. (A–E) COL X and DAPI are shown in blue in the merged images. (F–J) CD31 and DAPI are shown in green and blue, respectively, in the merged images. (K–O) Alizarin Red S staining histological images. All main and inset images are shown at 200x and 100x optical zoom, respectively. The scale bars are 100 μm.