Fig. 6

Study of tumor secretome profile and MDSC investigation on sarcoma cell lines. A NSG mice were engrafted with three different human sarcoma cell lines (WT, Crisped 14B OS Cell Lines for CXCL8 and G-CSF3). B Tumor growth was evaluated by In vivo imaging system (IVIS) for all three different orthotopically models (200′000 cell/mice). No tumor bearing mice have been used as control group. Weekly plasma samples were collected from each tumor-bearing mouse and control mice, to investigate circulating mMDSCs using FACS flow-cytometry. The analysis evaluated both monocytic MDSCs C and polymorphonuclear MDSCs D cell subset of CD11b + cells. To ensure accurate comparison and data normalization across samples, counting beads were utilized. *p-value = < 0.05; **p-value = < 0.01; ***p-value = < 0.001; ****p-value = < 0.0001, t test. E Quantification of the specified cytokines, including CCL2, CCL3, CCL5, CCL7, CX3CL1, CXCL1, CXCL2, CXCL4, CXCL5, CXCL6, CXCL8 (IL8), CXCL13, CXCL14, G-CSF (CSF3), GM-CSF, IFN-gamma, IL10, IL13, IL1b, IL5, MCSF, and TNF-alpha, was normalized on blank control background for the in vivo data, and single values reported in a Heatmap graph. Two-way Anova was used as multi comparison test. *p < 0.05, **p < 0.01, ***p < 0.001