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. 2022 Oct 23;13:6308. doi: 10.1038/s41467-022-34064-4

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© The Author(s) 2022

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 Immunohistochemistry analysis of CD47 expression (CD47 H-score) in matched osteosarcoma specimens from the same patient before and after chemotherapy (n = 81 patients). Scale bar, 400 μm. b, c Cisplatin or doxorubicin was injected intravenously every 2 days, and ifosfamide or methotrexate was injected intravenously every 4 days, after GFP⁺ HOS tumor grew for 14 days. b Western blot analysis of CD47 expression in HOS tumors on day 22. c Flow cytometric analysis of CD47 protein expression in GFP⁺ HOS cells from tumors on day 22 (left; n = 4 mice per group). The anti-CD47 median fluorescence intensity (MFI) was determined (right). d Kaplan–Meier curves of overall survival (OS) of osteosarcoma patients stratified by CD47 expression in pre-chemotherapy specimens. High CD47 expression (CD47^high): CD47 H-score > 0.9; low CD47 expression (CD47^low): CD47 H-score ≤ 0.9. e, f ΔCD47 H-score for each patient was obtained from subtracting CD47 H-score of pre-chemotherapy specimen from that of paired post-chemotherapy specimen. e Kaplan–Meier curves of overall survival (OS) of osteosarcoma patients stratified by ΔCD47 expression. High ΔCD47 expression (ΔCD47^high): ΔCD47 H-score > 0.8; low ΔCD47 expression (ΔCD47^low): ΔCD47 H-score ≤ 0.8. f Kaplan–Meier curves of overall survival (OS) of osteosarcoma patients stratified by CD47 expression in pre-chemotherapy specimens and ΔCD47 expression. Low CD47 expression and low ΔCD47 expression (CD47^low ΔCD47^low): CD47 H-score ≤ 0.9 and ΔCD47 H-score ≤ 0.8; low CD47 expression and high ΔCD47 expression (CD47^low ΔCD47^high): CD47 H-score ≤ 0.9 and ΔCD47 H-score > 0.8; high CD47 expression and low ΔCD47 expression (CD47^high ΔCD47^low): CD47 H-score > 0.9 and ΔCD47 H-score ≤ 0.8; high CD47 expression and high ΔCD47 expression (CD47^high ΔCD47^high): CD47 H-score > 0.9 and ΔCD47 H-score > 0.8. Data are shown as the mean ± SD. ns, not significant. *P < 0.05, **P < 0.01, ***P < 0.001, paired two-tailed Student t test (a), one-way ANOVA (c) or log-rank test (d–f). The experiment was performed three times with similar results (b, c). See the Source Data file for the exact P values. Source data are provided as a Source Data file.