Fig. 6: Effects of tumor-derived paracrine factors on TAMs and spheroids.

a Heat map representation of differentially expressed genes involved in macrophage recruitment, Wnt signaling, and cell adhesion in recovered peritoneal cells from ID8 bearing mice. b ID8 cells were cultured in DMEM overnight and the protein levels of CCL2 and CSF-1 in cell cultures were measured by ELISA (n = 5–6 biologically independent samples per group). c Western blotting showing the protein expression of UBR5/β-catenin/CCL2 in different ID8 cells. d Peritoneal macrophages were harvested and seeded into Transwell inserts with ID8 cell cultured in the bottom. Migrated macrophages were quantified after 16h-incubation (n = 5 biologically independent samples per group). e Representative images of spheroids. Scale bars: 200 μm. f Quantification of spheroid expansion rates (n = 4 biologically independent samples per group). g, h CD68⁺ cells and Ki67⁺ cells in spheroids were quantified (n = 4 mice per group). i Representative images of CD68⁺ macrophages and surrounded Ki67⁺ cells. (n = 4 mice per group and ten confocal images acquired from each sample). All panels are the same magnification, scale bars: 50 μm. j Spheroid retrieved from ID8 bearing mice at day 30 were underwent H&E staining (n = 4 mice per group and an average of five fields acquired from each sample). Scale bars: 200 μm. k Survival in mice bearing ID8 tumors (n = 5 mice per group), log-rank test. All data are representative of at least two independent experiments with similar results. Data are presented as mean ± SEM, unpaired two-sided Student’s t-test with no correction for multiple comparison, *P < 0.05, **P < 0.01, ***P < 0.001. Source data are provided as a Source Data file.