Fig. 3

CAFs-derived SCUBE1 can regulate the malignant progression of HCC cells.(A) PLC and Huh7 cells were cocultured with CAFs1, CAFs2 and CAFs3 with or without anti-SCUBE1 antibody. Cell viability was measured with CCK-8 assays (***P < 0.001 vs. CAFs2 or CAFs3, one-way ANOVA). (B) RT–qPCR was performed to determine the mRNA levels of CD133, Sox2 and Nanog in PLC and Huh7 cells cocultured with CAFs1 and CAFs2 (*P < 0.05, t test). (C) The expression levels of CD133, Sox2 and Nanog in treated and untreated HCC cells were evaluated by Western blotting (***P < 0.001, t test). (D) CD133 was detected via immunofluorescence staining in different treated HCC cells (**P < 0.01, t test). Scale bar, 10μm. (E) The effects of CAFs1, CAFs2 and anti-SCUBE1 antibody on the colony formation ability of PLC and Huh7 cells (*P < 0.05, **P < 0.01, ***P < 0.001 vs. CAFs2, one-way ANOVA). (F) HCC cell migration ability was evaluated with Transwell assays after coculture with CAFs1 and CAFs2 with or without an anti-SCUBE1 antibody (**P < 0.01, ***P < 0.001 vs. CAFs2, one-way ANOVA). Scale bar, 200μm. Data are represented as mean ± SD of three independent experiments