Fig. 7

circSMARCC1 promotes M2 macrophage polarization and recruitment via the CCL20-CCR6 axis. A IHC detected the infiltration of CD68⁺/ CD163⁺/ CD206⁺ macrophages in human PCa tissue and Para-cancerous tissues (scale bar, 100 μm and 20 μm). B IHC detected the infiltration of CD68⁺/ CD163⁺/ CD206⁺ macrophages in mouse xenograft tumors (scale bar, 100 μm and 20 μm). C, D The expression of M1 phenotype (TNF-a, CD80 and CD86) and M2 phenotype (IL-10, ARG-1 and CD163) markers were detected by qRT-PCR in THP-1 cells, THP-1-Mø and THP-1-M2 cells. E The proportion of CD68 and CD163 positive cells detected by flow cytometry. F The CM prepared from lv-circSMARCC1 or vector and sh-circSMARCC1 or sh-nc tumor cells was used to evaluate the migration ability of macrophages through transwell experiments (scale bar, 50 μm). G The migration ability of TAM was detected by transwell assay using CCL20 recombinant protein and CCR6 neutralizing antibody (scale bar, 50 μm). H The CM prepared from THP-1-Mø or THP-1-M2 cells for PCa cells migration experiments (scale bar, 50 μm). I, J qRT-PCR tested the expression of M2 phenotypes marker (IL-10, ARG-1, CD68 and CD163) when co-cultured with DU145-lv-circSMARCC1 cells or vector cells or using CCL20 recombinant protein, with IL-4 and IL-3 polarization as the reference. K The expression of CD68, CD163 and CCR6 in THP-1 cells, THP-1-Mø cells stimulated by CCL20 recombinant protein, and THP-1-Mø cells (with and without co-culture) were detected by Western blotting. L The expression of CD68, CD163 and CCR6 in THP-1-Mø cells and THP-1-M2 cells (with and without anti-CCR6) were detected by Western blotting. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001