Fig. 4

Effects of altered SCUBE1 expression in CAFs on the malignancy of HCC cells.(A) WB was used to detect the transfection efficiency of the overexpression plasmid and knockout plasmid. (B) RT–qPCR was used to detect the transfection efficiency of the overexpression plasmid (***P < 0.001 t test) and knockout plasmid (*P < 0.05, **P < 0.01, ***P < 0.001 vs. sh-Control, one-way ANOVA). (C) Cell viability was measured with CCK-8 assays in HCC cells subjected to different treatments (***P < 0.001, t test). (D) The effect of SCUBE1 silencing and overexpression in CAFs on the migration of HCC cells was measured with Transwell assays (**P < 0.01, t test). Scale bar, 200μm. (E) Sphere formation ability was detected in PLC cells and Huh7 cells. Scale bar, 500μm. (F) Shh, Smo, and Gli1 protein expression levels were detected via Western blotting in PLC and Huh7 cells cocultured with different treated CAFs. (G) Growth curve of mouse subcutaneous tumours derived from different treated PLC cells at the indicated time points (***P < 0.001, t test). Data are represented as mean ± SD of three independent experiments