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. 2021 Jul 2;12:683939. doi: 10.3389/fgene.2021.683939

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Copyright © 2021 Bian, Zhu, Liang, Hei, Zhang, Li, Chen, Lu, Gu, Qiao and Zheng.

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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Transfection of Tbx5 in MCT cells. (A) Schema graph of Tbx5 expression plasmid driven by Col10a1-specific enhancer and promoter. (B) The Col10a1 gene (top) and Col10a1–Tbx5 transgenic construct (bottom). The ~300-bp hypertrophic chondrocyte-specific Col10a1 cis-element illustrated previously locate in the distal promoter (−4,296 to −4,009 bp) and four copies of the ~300-bp cis-elements and a short basal promoter (265bp) element were used to drive the Tbx5 gene with a Flag-tag. ATG: start codon; TAG: stop codon; ShXBP: Short Col10a1 basal promoter. (C,D) Transient transfection of Tbx5 increased the mRNA level of Tbx5 and downregulated Col10a1 expression. (E,F) Compared with scrambled RNA, Tbx5 siRNA downregulated Tbx5 expression and increased Col10a1 expression. *p < 0.05, **p < 0.01.