Fig. 6

Tumor-associated macrophages show higher immunosuppressive capacity in the hybrid cell-induced microenvironment. a Representative tSNE plots of bone marrow from normal mice (left), mice with RM1-derived bone metastasis (middle), and mice with hybrid cell-derived bone metastasis (right). b The percentage of myeloid cells (left) and B cells (middle) in CD45⁺ cells, the percentage of macrophages (right) in CD45⁺ CD11b.⁺ cells (n = 4). c–e Representative CyTOF dot plots showing the percentage of B cells (c), myeloid cells (d), and macrophages (e) in different groups. f Schematic illustration of macrophages cocultured with RM1 or hybrid cells. g qRT–PCR for Arg-1, Ccl2, Cd206, Il-11, Tgfb, and Ptgs2 expression in macrophages cocultured with RM1 or hybrid cells (n = 4). h Heatmap showing the expression of chemokines and cytokines related to myeloid cell recruitment and differentiation in RM1 cells and hybrid cells. i Cytokine array analysis of cell culture supernatant from RM1 cells and hybrid cells. j GO enrichment analysis of up-regulated cytokines (Log₂(fold change) > 1) in hybrid cells, based on the result of cytokine array. k High expression of Ccl2, Ccl3, Ccl5, Csf1, Csf2, and Csf3 in hybrid cells can recruit myeloid cells to the tumor, and then induce them to polarize into N2-like neutrophils or M2-like macrophages to inhibit the immune response. ns: not significant, *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001