Fig. 1

CRC promoted HSCs-to-CAFs differentiation by releasing VEGF. A Figure demonstrates the experimental set up for control (upper panel) and experiment (lower panel) for the conditioned medium (CM) system. B Cell morphology of LX-2 after treated with LX-2-CM, HCT-116-CM or LOVO-CM medium. C Effects of HCT-116-CM or LOVO-CM on the protein levels of FAP and α-SMA. Protein levels were examined using immunoblotting (left panel); and quantitative results were analyzed using Image J software (right panel). D Cell morphology of JS1 after treated with JS1-cultured medium or CT26-cultured medium. E Effects of CT26-CM on the protein levels of FAP and α-SMA. Protein levels were examined using immunoblotting (left panel); and quantitative results were analyzed using Image J software (right panel). F The levels of the cytokines or growth factors in the HCT-116 conditioned medium were detected by using the Luminex liquid suspension chip assay. G The VEGF contents in the medium of NCM460, HCT-116 or LOVO cells were detected by using the ELISA assay, and the VEGF contents in the medium of HCT-116 or LOVO cells treated with neutrakine VEGF165/121 monoclonal antibody were also detected by using ELISA assay. H Effects of VEGFA on the protein levels of VEGF, FAP and α-SMA. Protein levels were examined using immunoblotting (left panel); and quantitative results were analyzed using Image J software (right panel). I Effects of HCT-116-CM and HCT-116-CM + VEGF neutralizing antibody on the protein levels of FAP and α-SMA. J Effects of LOVO-CM and LOVO-CM + VEGF neutralizing antibody on the protein levels of FAP and α-SMA. Protein levels were examined using immunoblotting (left panel); and quantitative results were analyzed using Image J software (right panel). Data are shown as mean ± SD from three independent experiments, for G, **P < 0.01, *P < 0.05 vs. NCM460; ^##P < 0.01 vs. CRC cells; for C, E and H, **P < 0.01, *P < 0.05 vs. the corresponding control. for I and J, **P < 0.01 vs. HSCs-CM, ^#P < 0.05, ^##P < 0.01 vs. CRC-CM.CM: cultured medium