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. 2023 Nov 30;42:328. doi: 10.1186/s13046-023-02886-9

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 5 — Proteomic analysis identified EFEMP1 as a potential metastatic-related biomarker in osteosarcoma. A, B Gene ontology analysis (GO) output. Biological process (BP) and molecular function (MF) of differently expressed proteins (DEPs) in the two pairwise-comparison groups (SCR 143B vs. MG-63; Plasma PT vs. CTR). Circle sizes in BP denote the number of genes involved in the process. C Reactome pathway enrichment analysis of identified proteins in the two pairwise- comparison groups. Circle sizes denote the number of genes included in a group and colour indicates the p-value. The common pathways are highlighted. D Venn diagram showing specific and common proteins among the two pairwise groups: SCR 143B vs. MG-63 and Plasma PT vs. CTR. E EFEMP1 levels in the SCR of the metastatic 143B and non-metastatic MG-63 OS cells. F Representative western blot of EFEMP1 in the metastatic 143B and non-metastatic MG-63 OS cells. G Plasma levels of EFEMP1 in control mice (CTR), mice treated with the 143B SCR, bearing a primary tumour (PT) or with lung metastasis (Lung Met). H Representative western blot of EFEMP1 expression in 143B cells (CTR), non-targeting (NT) and siRNA knockdown of EFEMP1 cells. I EFEMP1 levels in the SCR of 143B cells, NT and siRNA EFEMP1-knockdown cells. J Representative bioluminescence images of mice treated with the SCR of control 143B (SCR + i.v. group) or EFEMP1-knockdown cells (SCR siRNA EFEMP1 + i.v. group) followed i.v. injection of 143B-Luc⁺ cells. Data are presented as mean ± SEM, from 7 to 8 independent experiments. **** p < 0.0001 were significantly different when compared with the SCR from MG-63 (unpaired t-test (E)); *p < 0.05, **p < 0.01 and ***p < 0.001 were significantly different when compared with the values present in the plasma from healthy mice (Kruskal-Wallis test (G); *p < 0.05, **p < 0.01 were significantly different when compared with 143B SCR (Kruskal-Wallis test (I))