Fig. 5.

M2-Mφ synergized the malignant phenotype of TNBC via PPP. A The macrophages markers CD206, IL-10, IL-12, IL-1β and IFNγ were determined using q-PCR. B Protein expression in macrophages was examined using immunoblotting. C G6PD enzyme activity was detected in macrophages. D M0-Mφ and M2-Mφ were treated with different concentrations of 6-AN (0, 1.2, 2.4, 5, 10, 20, 40, 78, 156, 313 and 625 μM) for 48 h. Cell viability after drug treatment was measured using the MTT assay. E Altered proteins in M0-Mφ incubated with CM were measured by western blotting. F G6PD enzyme activity was detected in macrophages. G MDA-231 cells and macrophages were cocultured in the transwell migration experiment. Magnification 40 × , bar = 200 μm. H The cell viability of MDA-231 cells cocultured with CM from macrophages for 48 h detected by MTT assay. I The proliferation of MDA-231 cells cocultured with CM from macrophages for 48 h detected by the EdU assay. The green color represents EdU positivity, and the blue color represents DAPI (nucleus). Representative images at 630 × magnification is presented, bar = 25 μm. J G6PD enzyme activity was detected in MDA-231 cells coincubated with CM from macrophages. K Altered protein in MDA-231 cells incubated with the supernatant medium of macrophages was measured by western blot. H–K CM was transferred to MDA-231 cells after mixing 1:1 with fresh media. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001