Fig. 3

XIAOPI formula inhibits the proliferation and invasion of 4 T1 cells co-cultured with TAMs and HSPCs. a EdU cell proliferation assay demonstrated the co-culture of HSPCs and TAMs-CM promoted the proliferation of 4 T1, while was inhibited by XIAOPI formula or CXCL1 neutralizing antibody (the scale bars indicate 50 μm). b The invasion of 4 T1 cells was significantly promoted by co-culture of HSPCs and TAMs-CM, while was inhibited by XIAOPI formula treatment and CXCL1 neutralizing antibody. Meanwhile, the suppressive effects of XIAOPI was reversed by exogenous CXCL1 treatment (the scale bars indicate 200 μm). c Co-culture with HSPCs and TAMs-CM significantly promoted the expression of MMP2 and MMP9, while XIAOPI formula or CXCL1 neutralizing antibody reduced the elevated levels of MMP2 and MMP9. However, exogenous CXCL1 could partly restore MMP2 and MMP9 levels that were suppressed by XIAOPI formula. d Quantification of fluorescence intensity in 4 T1 cells (per cell) indicated that XIAOPI formula and CXCL1 neutralizing antibody inhibited the mesenchymal marker vimentin, which was upregulated by co-culture with HSPCs and TAMs-CM. By contrast, the expression of epithelial marker E-cadherin was enhanced (the scale bars indicate 20 μm). (The results were obtained from triplicate experiments and were represented as mean values ± SD, *P < 0.05, **P < 0.01)