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. 2023 Apr 25;14:2383. doi: 10.1038/s41467-023-38034-2

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© The Author(s) 2023

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. 9 — a CFU-F assay of Fgfr3-p53 cHet and Fgfr3-p53 cKO bone marrow cells at P28 (pulsed at P21). Scale bar: 5 mm. n = 4 mice per each group. b Survival curve of individual tdTomato⁺ clones over serial passages among Fgfr3-WT (P23, red), Fgfr3-p53 cKO (P28, blue), and Fgfr3-p53 cKO (P4M, black). n = 36 (Fgfr3-WT P23), n = 35 (Fgfr3-p53 cKO P28), n = 18 (Fgfr3-p53 cKO P4M) clones. Log-rank (Mantel–Cox) test. c Duration to reach Passage 8. n = 5 (Fgfr3-WT P23), n = 29 (Fgfr3-p53 cKO P28), n = 12 (Fgfr3-p53 cKO P4M) clones. Two-tailed, one-way ANOVA followed by Tukey’s multiple comparison test. d, e Comparative bulk RNA-seq analysis of individual tdTomato⁺ clones, Fgfr3-WT (P23, n = 5 clones), Fgfr3-p53 cKO (P28, n = 5 clones) and Fgfr3-p53 cKO (P4M, n = 4 clones) clones. d Principal component analysis (PCA) plot of Fgfr3-WT (P23, light green), Fgfr3-p53 cKO (P28, blue) and Fgfr3-p53 cKO (P4M, light blue) clones. x-axis: PC1, 37.2% variance, y-axis: PC2, 14.2% variance. e Heatmap of representative differentially expressed genes (DEGs) associated with osteoblast differentiation, cell proliferation, and osteosarcomagenesis. Star: statistically significant difference among the three groups. n = 4 (Fgfr3-p53 cKO P4M), n = 5 (Fgfr3-WT P23, Fgfr3-p53 cKO P28) mice. A two-sided t-test, based on a beta-negative binomial distribution, was used to calculate the p values (corrected for multiple testing) with an FDR cutoff of 0.05 to identify DE for the bulk RNA-seq data. f Diagram of intrafemoral transplantation of individual Fgfr3-WT (P23) and Fgfr3-p53 cKO (P28) tdTomato⁺ clones to NSG recipient mice. g 3D-μCT of Fgfr3-WT (P23) and Fgfr3-p53 cKO (P28) transplanted femurs after 8 weeks of transplantation. Scale bar: 500 µm. n = 4 per each group. h Histology of recipient femurs. (Left): Transplantation of Fgfr3-WT (P23) clones after 8 weeks of transplantation. (Left center): Transplantation of Fgfr3-p53 cKO (P28) clones after 4 weeks (left center) and 8 weeks (right center) of transplantation. (Right): Magnified views of highly-trabecularized areas in recipient bone marrow, fluorescent, and H&E staining (right). Scale bar: 500 µm. n = 4 per each group. Data were presented as mean ± s.d. Exact P value is indicated in the figures. Source data are provided as a Source Data file.