Fig. 4

M2 TAMs derived PDGF-BB promotes tumor angiogenesis in vitro. A Gene set enrichment analysis in COAD conducted on the TCGA database. B Correlation analysis between RUNX1 and endothelial cell markers in CRC tissue samples. C Correlation analysis between RUNX1 and PDGFB (left), PDGFRA (middle) and PDGFRB (right) in the TCGA database. D Wikipathway cancer over-respresentation analysis of RUNX1 associated pathways. E The expression of PDGF-BB in THP-1 treated with CMs derived from RUNX1 over-expressed HCT116 and RKO cells was determined by ELISA. F The expression of PDGF-BB in THP-1 treated with CMs derived from RUNX1 down-regulated SW480 and RKO cells was determined by ELISA. G The expression of PDGF-BB in HCT116 RUNX1^OE/THP-1 co-culture supernatant was determined by ELISA. H The expression of PDGF-BB in SW480 shRUNX1/THP-1 co-culture supernatant was determined by ELISA. I The 48 h proliferation rate of HUVECs treated with HCT116 RUNX1^OE/THP-1 co-culture supernatant with or without anti-hPDGF-BB antibody (20 ng/ml) was detected. J (left) Transwell migration assay. The ability of HCT116 RUNX1^OE/THP-1 co-culture supernatant to promote HUVECs migration in the presence or absence of anti-hPDGF-BB antibody (20 ng/ml) was evaluated. Scale bars: 200 μm. (right) Quantification of the number of cells migrating to the lower chamber. K (left) Transwell invasion assay. The ability of HCT116 RUNX1^OE/THP-1 co-culture supernatant to promote HUVECs invasion in the presence or absence of anti-hPDGF-BB antibody (20 ng/ml) was evaluated. Scale bars: 200 μm. (right) Quantification of the number of cells invading to the lower chamber. L (left) Tube formation on Matrigel. The ability of HCT116 RUNX1^OE/THP-1 co-culture supernatant to promote HUVECs tube formation in the presence or absence of anti-hPDGF-BB antibody (20 ng/ml) was evaluated. Scale bars: 200 μm. (right) Quantification of the branch points of HUVECs. M (left) Transwell migration assay. The ability of PDGF-BB to promote HUVECs migration was evaluated. Scale bars: 200 μm. (right) Quantification of the number of cells migrating to the lower chamber. N (left) Transwell invasion assay. The ability of PDGF-BB to promote HUVECs invasion was evaluated. Scale bars: 200 μm. (right) Quantification of number of cells invading to the lower well. O (left) Tube formation on Matrigel. The ability of PDGF-BB to promote HUVECs tube formation was examined. Scale bars: 200 μm. (right) Quantification of the branch points of HUVECs