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. 2020 Jan 13;10:157. doi: 10.1038/s41598-019-57011-8

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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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LacZ expression is upregulated during cartilage repair and in vitro chondrogenesis. (A) Areas of healed cartilage (dashed line) in the patellar groove of the femur of Gdf5DOWN-LacZ (n = 4/10) and Gdf5UP-LacZ (n = 3/10) mice, with LacZ-expressing chondrocytes (blue, arrows) detected in Gdf5DOWN-LacZ mice 4 weeks post-injury. LacZ, whole-mount X-gal staining to detect LacZ expression; SafO & FG, Safranin O and Fast Green counterstaining; Fast Red, Nuclear Fast Red counterstaining. Scale bars, 100 μm. (B) Histological sections of chondrogenic cell pellets. Synovial cells were isolated from Gdf5DOWN-LacZ and Gdf5UP-LacZ mice and treated in vitro for 21 days with TGFβ (10 ng/ml) or BMP-2 (300 ng/ml) to induce chondrogenesis, followed by X-gal staining to detect LacZ expression. Tol blue, Toluidine blue metachromatic staining indicates deposition of cartilage proteoglycans; Fast Red, Nuclear Fast Red counterstaining. LacZ-expressing chondrocytes (blue, arrows) were observed in Gdf5DOWN-LacZ cell pellets, but not Gdf5UP-LacZ pellets, under both culture conditions. Scale bars, 100 μm.