Fig. 2

Enhancement of stemness in HCC cells induced by myofibroblasts was abolished after the knockdown of ITGA5. A Volcano plot showing the differentially expressed genes (DEGs) detected by RNA-sequencing (RNA-seq) in Huh7 cells cultured with conditioned medium (CM) from myofibroblasts (TGF-β1 induced LX-2 cells) with or without ITGA5 knockdown (si-Ctrl CM vs. si-ITGA5 CM). B GO enrichment analysis of DEGs. C The GSEA revealed several signaling pathways that were notably enriched among the DEGs. D Sphere formation assays were conducted in Huh7 cells with CM of myofibroblasts (si-Ctrl or si-ITGA5). Scale bar: 50 μm. E Schematic of the subcutaneous xenograft mouse model injected with Huh7 cells alone or co-injected with myofibroblasts (vector or ITGA5-KO). F Variation in tumor volume among the indicated groups. G Quantification of the tumor weights in the indicated groups (n = 6 for each group). H Respective IHC staining images and quantification of ITGA5 and EPCAM in the tumor tissues of the indicated groups. Scale bar: 50 μm. I Representative immunofluorescence images of ITGA5 (red), α-SMA (green), and ALDH (pink) in human normal liver tissue or HCC tissues. Scale bar: 50 μm. *P < 0.05; **P < 0.01; ***P < 0.001. Data were obtained from three independent experiments per group