Fig. 6

M2-TAMs derived PDGF-BB promotes malignant biological behavior of CRC cells A The levels of PDGF-BB in THP-1 culture medium with or without IL-4 stimulation were detected by ELISA. B The RUNX1 expression in HCT116 and SW480 cells treated with M0 or M2 TAMs culture supernatant in the presence or absence of anti-hPDGF-BB antibody were detected by western blotting. C RT-qPCR analysis of the RUNX1 mRNA expression in CRC cells treated with M0 or M2 TAMs culture supernatant. D (left) Transwell invasion assay. The invasion rate of HCT116 and SW480 cells treated with M0 or M2 TAMs culture supernatant were evaluated. Scale bars: 200 μm. (right) Quantification of the number of cells invading to the lower well. E (left) Transwell migration assay. The migration rate of HCT116 and SW480 cells treated with M0 or M2 TAMs culture supernatant were evaluated. Scale bars: 200 μm. (right) Quantification of the number of cells migrating to the lower chamber. F Wound healing assay. The ability of the M0 or M2 TAMs culture supernatant to promote CRC cells wound healing was evaluated. G The changes of RUNX1 expression in the cytosol and nuclei of HCT116 and SW480 cells upon the M0 or M2 TAMs culture supernatant stimulation were assessed by western blotting. H The changes of RUNX1 expression in the cytosol and nuclei of CRC cells treated with PBS or 20 ng/ml PDGF-BB were detected by western blotting. I, J RUNX1 IF labeling of HCT116 or SW480 cells treated with M0 or M2 TAMs culture supernatant. Data were recorded by confocal laser scanning. Scale bars: 10 μm. K The protein expression of c-myc, LEF1, cyclinD1 and GAPDH in HCT116 and SW480 cells treated with PBS or 20 ng/ml PDGF-BB were detected by western blotting. L The pulmonary metastasis model of CRC in nude mice. (left, middle) The full image of metastatic tumors. HCT116 cells were firstly stimulated with THP-1-derived CMs for 24h, and then inoculated into nude mice via caudal vein. (right) Quantification of numbers of the lung tumor nodules in different groups (n = 5). M Representative H&E staining analysis of the lung tumor nodules (#3, #4). Scale bars: 100 μm