Fig. 5

HCC cell-derived SPP1 enhanced LX2’s migratory ability and upregulated CAF-specific markers. (A): Tumor cell-derived CM stimulated LX2 to express CAF-specific markers (n=3). (B-C): 97 h and huh7 cells were infected with SPP1 shRNA lentiviral to silence SPP1 expression, and knockdown efficiency was verified by WB (n=3). (D): A flow chart illustrated the culture process of tumor cell CM against LX2. (E): SPP1 from cancer cells increased LX2 migration, demonstrated by a transwell assay after LX2 was cultured with CM from control or SPP1 knockdown tumor cells for 48 h (n=3). (F): Cancer cell-derived SPP1 enhanced LX2 proliferation (n=3). G-H: HCC cell-derived SPP1 upregulated CAF-specific markers, as validated by CM from 97 h (G) and huh7 (H) before and after SPP1 knockdown (n=3). (I): Immunohistochemistry confirmed SMA expression differences between tumor and paracancerous tissues, showing a positive correlation between IHC scores of SPP1 and SMA in tumor tissues (n=20). *p<0.05, **p<0.01, ns: p>0.05