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. 2020 Feb 20;10:3097. doi: 10.1038/s41598-020-60047-w

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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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Comparison of basal levels in cells derived from middle and distal third of ACLR regions. (a) Cell proliferation rates were determined using a WST assay. X axis: days; Y axis: OD value (450 nm). Red, Middle (n = 5); Blue, Distal (n = 5). (b) mRNA expressions of ALP, RUNX2, SOX9, and COL2 were compared by qRT-PCR. Red, Middle (n = 6); Blue, Distal (n = 6). (c) Expressions of ALP and SOX9 in both middle and distal cells were compared by immunofluorescence. Green, ALP-alexa-488; Red, SOX9-cy3; Blue, DAPI; Scale bar: 200 μm. Representative image data are shown (n = 3). (d) Expression of RUNX2, SOX9, ALP, and GAPDH were detected by immunoblotting (n = 5). (e) Quantification of (d). Red, Middle (n = 5); Blue, Distal (n = 5). Error bars show standard error of the mean. *p < 0.05. **p < 0.01.