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. 2021 Mar 17;12:1706. doi: 10.1038/s41467-021-21948-0

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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. 1 — a Immunohistological staining (first row, pSmad2/3: brown), immunofluorescence staining (second row, pSmad2/3: green, DAPI: blue) staining of pSmad2/3 in AC, and representative µCT images of SB (third row) of human knee joint specimens. HC healthy control, OA-M OA specimen with minimal cartilage degeneration, OA-S OA specimen with severe cartilage degeneration. b Quantitative analysis of the ratio of pSmad2/3⁺ cells in total chondrocytes in AC, n = 5 or 25 biologically independent joint samples in HC or OA-M and OA-S group, respectively. c–e Quantitative analysis of µCT scanning: BV/TV: percent bone volume, BS/BV: bone surface/volume ratio, SMI: structure model index, n = 6, 10, and 9 biologically independent joint samples in HC, OA-M or OA-S group, respectively. In (b–e), data were analyzed using one-way ANOVA followed by LSD post hoc test. Data are presented as mean values + /− SEM. Source data are provided as a Source data file. f, g The region-defining strategy for medial compartment of mouse tibia. f Top view of tibia plateau for illustration. The gold dotted line represents the location of the sagittal images in g, the black dotted lines represent the locations of the coronal images in h. P posterior, M middle, A anterior. h Representative 3D reconstructed µCT images of coronal sections of the medial compartment of mice tibia (first and fourth columns), and immunohistological staining of pSmad2/3 (brown) in AC at the corresponding locations (second, third, fifth, and sixth columns). The third and sixth columns are the high-magnification images of the box region in columns 2 and 5, respectively. The mice were sacrificed 2 months after sham surgery or ACL-T. i The ratio of pSmad2/3⁺ cells to total chondrocytes in different regions of AC in h. Data were analyzed using two-way ANOVA Tukey’s post hoc test. j The correlation of trabecular pattern factor (Tb. Pf) of the SB and the percentage of pSmad2/3⁺ cells in the above cartilage at the same location. h–j n = 6 biologically independent animals, data are presented as mean values + /− SEM.