Fig 1: Kaplan-Meier analysis of DFS based on TAMs and macrophage-related biomarkers. (A–D) Kaplan-Meier analysis of DFS based on pre-NCRT CD163, CD68, MCSF, CCL2 scores. (E–H) Kaplan–Meier analysis of DFS based on post-surgical CD163, CD68, MCSF, CCL2 scores.
Fig 2: Association of pre-NCRT TAMs and macrophage-related biomarkers with pCR. (A) Heat map of pre-NCRT TAM scores, macrophage-related biomarker levels and clinical parameters. The left Y-axis were the CD163 and CD68 scores and the right Y-axis were the MCSF and CCL2 scores. (B–E) Pre-NCRT TAMs were lower in the pCR group when compared with the non-pCR group (all P<0.01). (F–I) ROC analysis demonstrating that pre-NCRT CD163 (AUC=0.77, P<0.01), pre-NCRT CD68 (AUC=0.66, P<0.01), pre-NCRT MCSF (AUC=0.66, P<0.01), and pre-NCRT CCL2 (AUC=0.67, P<0.01) scores had a powerful ability to predict pCR.
Fig 3: Kaplan–Meier analysis of OS based on TAMs and macrophage-related biomarkers. (A–D) Kaplan–Meier analysis of OS based on pre-NCRT CD163, CD68, MCSF, CCL2 scores. (E–H) Kaplan–Meier analysis of OS based on post-surgical CD163, CD68, MCSF, CCL2 scores.
Fig 4: Construction of miRNA-mRNA network. (A–E) Overlap analysis for the miRNAs binding to the IL6, Jun, Cd44, Timp1, and Csf1 from the TargetScan, miRWalk, and miRDB database. (F) The miRNAs-hub genes network.
Fig 5: MYC association with CSF1 expression.(A) Western blot of CSF1 protein expression in Myc-knockin tumors compared with the p53-driven tumor. The quantified expression is shown in the right panel. (B) The RNA-Seq analyzed relative mRNA expression, demonstrating reduced Csf1 mRNA expression in Myc-knockin (n = 5) tumors compared with the p53-driven (n = 4) tumor samples. (C) IHC staining for CSF1 in paraffin-embedded GEMM tumor tissue samples in the Myc-knockin tumors (lower panel) compared with p53-driven tumor (upper panel). The quantified expression is shown in the right panel. (D) MYC and CSF1 mRNA expression in the PDX samples of the OS. (E) Negative correlation between the MYC and CSF1 mRNA expression in human OS TARGET data set patients. (F) Kaplan-Meier curve of human OS TARGET data set for CSF1 expression with top quartile (n = 22) or bottom quartile (n = 22) samples; log-rank (Mantel-Cox) test was performed for the Kaplan-Meier analyses. (G) Csf1 expression upon transient knockdown of Myc in Myc-knockin murine OS cell lines. (H) Csf1 mRNA expression after dTAG-13 and -v1 treatment. (I) Western blot of MYC and CSF1 protein expression after dTAG-13 and -v1 treatment; blot quantification is shown in the right panel. (*P < 0.05, **P < 0.01, ****P < 0.0001.)
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