Fig. 1

HCC malignancy was promoted after cocultivation.(A) The expression of α-SMA in LX2 cells and cocultured LX2 cells was detected by WB. (B) Cell viability was evaluated with CCK-8 assays of PLC and Huh7 cells cocultured with LX2 cells (**P < 0.01, ***P < 0.001, t test). (C) The colony formation ability of PLC and Huh7 cells cocultured with activated LX2 cells was detected with colony formation assays (*P < 0.05, ***P < 0.001, t test). (D) The migration ability of HCC cells and cocultured HCC cells was compared using Transwell assays (**P < 0.01, t test). Scale bar, 200μm (E) Immunofluorescence was used to assess CD133 protein expression in HCC cells and cocultured HCC cells (*P < 0.05, **P < 0.01, t test). Scale bar, 10μm. CD133 fluorescence density values in HCC cells were quantified before and after coculture. (F) Western blotting was used to assess CD133 protein expression in HCC cells and cocultured HCC cells (**P < 0.01, ***P < 0.001, t test). (G)CD133 mRNA expression was detected by RT–qPCR (**P < 0.01, ***P < 0.001, t test). (H) Sphere formation assays were used to evaluate the self-renewal capacity of treated and untreated HCC cells. Scale bar, 500μm. Data are represented as mean ± SD of three independent experiments