Research Paper Volume 15, Issue 6 pp 2321—2346

Downregulated annexin A1 expression correlates with poor prognosis, metastasis, and immunosuppressive microenvironment in Ewing’s sarcoma

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Figure 10. The expression levels of ANXA1-related module genes in ES cell lines and MSCs by RT-qPCR and Western blot and functional validation of ANXA1 in RD-ES cell using CCK-8 and EdU assays. The relative mRNA levels of ANXA1, MMP9, COL1A2, S100A4, VIM and S100A11 in RD-ES (A) or A673 (B) cells compared to MSCs. (C) The protein levels of ANXA1-related module genes in ES cell lines and MSCs by Western blot. (D) Quantitative analysis of protein expression levels of module genes. (E) The efficiency of si-ANXA1 was detected by RT-qPCR and Western blot analysis. (F) ES-RD cell viability at 24 h, 48 h and 72 h after downregulation of ANXA1 was measured using a CCK-8 assay. (G) The effect of ANXA1 on ES-RD cell proliferation was assessed by an EdU assay. P values are shown as follows: * P<0.05, ** P<0.01, *** P<0.001, and **** P<0.0001.