Fig. 4. RAD18-driven YAP activation in TNBC cells promotes M2-like TAM polarization.
A The Timer software showed that high-expressed RAD18 was correlated with high-level macrophage infiltration in BRCA. RAD18 expression level was proportional to M2-like TAM infiltration, whereas inversely proportional to M1-like TAM infiltration. B Schematic model of the co-culture system of macrophages and TNBC cells; THP-1 cells were stimulated by PMA for 48 h to form M0 macrophages, then MDA-MB-231 cells stably expressing shNC or shRAD18 were co-cultured with M0 macrophages for 72 h. The M0 alone were used as a negative control. C Detection of the expression of CD86 (M1-like macrophage marker) and CD163 (M2-like macrophage marker) in co-cultured macrophages by Flow cytometry analysis. D WB detection of protein expression of CD86 and CD163 in macrophages after co-culture. E The expressions of CD86 (red) and CD163 (green) in macrophages were detected using immunofluorescence staining (400×). Cell nuclei were stained with DAPI (blue). F The expression of CD86 and CD163 in macrophages co-cultured with the shNC TNBC treated with verteporfin or the shRAD18 TNBC treated with YAPOE plasmid were detected by flow cytometry analysis. *p < 0.05, **p < 0.01, ***p < 0.001. All experiments were performed independently at least three times.