Fig. 4

Brevilin A targets the VEGF-IL6-STAT3 axis in the CRC-HSCs interplay. A, B Brevilin A decreased viability in CRC (A) and NCM460 (B) cells. Cell viabilities were determined by the MTT assay. Data are shown as mean ± SD from three independent experiments, *P < 0.05, **P < 0.01 vs. vehicle (24 h); ^#P < 0.05, ^##P < 0.01 vs. vehicle (48 h). C The VEGF content in the medium of HCT-116 after treated with brevilin A were detected by using the ELISA assay. D, E, F Effects of brevilin A on protein levels of FAP and α-SMA in HCT-116-CM treated LX-2 cells (D), CT26-CM treated JS1 cells (E), or VEGFA-treated LX-2 cells (F) were determined by using Western blotting (left panel); and quantitative results were analyzed using Image J software (right panel). G, H The IL6 content in the medium of HCT-116-CM cultured LX-2 cells (G) or CT26-CM cultured JS1 cells (H) after treated with brevilin A were detected by using the ELISA assay. I Representative photographs of cell migration in HCT-116, HCT-116-LX-2 co-culture or brevilin A (1.25 μM and 2.5 μM)-treated co-culture system. J, K Effects of brevilin A on protein levels of STAT3, p-STAT3 (Tyr705), JAK2, p-JAK2 (Tyr1007/1008), SRC and p-SRC (Tyr416) in co-culture system were determined by using Western blotting (left panel); and quantitative results were analyzed using Image J software (right panel). Data are shown as mean ± SD from three independent expriments. For D, E, G, H: ^**P < 0.01, ^*P < 0.05 vs. HSCs; ^##P < 0.01, ^#P < 0.05 vs. CRC-CM treated HSCs. For F: ^**P < 0.01, ^*P < 0.05 vs. HSCs. ^##P < 0.01, ^#P < 0.05 vs. VEGFA-treated HSCs. For J, K: ^**P < 0.01, ^*P＜0.05 vs. CRC cells only; ^##P < 0.01, ^#P < 0.05 vs. co-culture with HSCs