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. 2021 Jul 19;12:4391. doi: 10.1038/s41467-021-24643-2

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© The Author(s) 2021

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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Fig. 5 — a–f Representative confocal images of repaired Achilles tendon immunostained with FGFR3 (a), BMPR1a (c) or pSmad1/5 (e) (white), LYVE1 (green), and DAPI (blue) in Col2^tomato (left) and FGFR3^f/f-Col2^tomato mice (right) at 8 weeks after surgery and relative quantitative analysis (b, d, f). n = 4 per group. Dashed line boxes indicate tdTomato-labeled LYVE1⁺ LECs (higher magnification with split channels, right). Scale bars, 100 μm (left); 20 μm (right). g Protein levels of FGFR3, BMPR1a, pSmad1/5, and Smad1/5 in mouse LEC cell line treated with FGFR3-targeted siRNA (KD) or control siRNA (CON). n = 3 per group. h–m Representative μCT (h), SOFG staining (j), and OC immunohistochemistry (l) images of ectopic bone in the Achilles tendon of FGFR3^Col2 (n = 4), FGFR3^Col2;BMPR1a^f/+ (n = 5), FGFR3^Col2;BMPR1a^f/f (n = 5), FGFR3^Prox1 (n = 4), FGFR3^Prox1;BMPR1a^f/+ (n = 10), and FGFR3^Prox1;BMPR1a^f/f mice (n = 6) at 8 weeks after surgery and relative quantitative analysis (i, k, m). Scale bars, 1 mm for μCT; 200 μm for SOFG and OC IHC. n–q Representative confocal images of Achilles tendon immunostained with LYVE1 (green) and DAPI (blue) (n) in FGFR3^Col2 (n = 4), FGFR3^Col2;BMPR1a^f/+ (n = 5), FGFR3^Col2;BMPR1a^f/f (n = 5), FGFR3^Prox1 (n = 4), FGFR3^Prox1;BMPR1a^f/+ (n = 5), and FGFR3^Prox1;BMPR1a^f/f mice (n = 6) and relative quantitative analysis (o, p) at 8 weeks after surgery. Scale bars, 100 μm. F4/80 (red), iNOS (green) and DAPI (blue) immunostained Achilles tendon sections (q). Scale bars, 20 μm. White dashed lines indicate outlines of the tendon (a, c, e, n). All data represent mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001 by unpaired two-tailed Student’s t-test (b, d, f) or by one-way ANOVA followed by a Tukey’s multiple comparisons test (i, k, m, o, p).