Fig. 3

CAF-derived STC1 promoted HCC stemness in a Notch1-dependent manner. A, B GSEA (A) and GO analysis (B) of the HCC samples in the TCGA database were performed. C Immunofluorescence images of MHCC-97H cells cultured with DMEM or rhSTC1 (20 ng/ml) to assess the localization of Notch1/NICD (green), Scale bars, 50 μm. D, E The protein levels of Notch1, NICD, HES1, HEY1, and STC1 were measured by western blotting. β-Actin was used as the loading control. F A sphere formation assay was used to determine the self-renewal ability of MHCC-97H and SNU-398 cells with Notch1 knockdown in the presence of DMEM or rhSTC1 (20 ng/ml). Scale bar, 50 μm. G After treatment with sorafenib for 24 h, the cell viability of different groups was assessed by a CCK8 toxicity assay. H The proliferation ability of different groups of HCC cells was evaluated by colony formation assay. I A Transwell migration assay was performed to detect the migration ability of the indicated MHCC-97H cells. Scale bar, 100 μm. J SNU-398-LUC cells with knockdown of Notch1 were mixed with stable CAF-STC1 cells lines and implanted in the livers of nude mice. The bioluminescence intensity of the groups was evaluated. For the statistical analysis, ns, no significance, *P < 0.05, **P < 0.01, and ***P < 0.001, t test