Fig. 5

HERC2 promoted the stemness and immune evasion of HCC cells through JAK2/STAT3 signaling. A HERC2 knockout Huh7 cells were treated with 50 ng/ml IL-6 for 24 h and then subjected to RNA-seq analysis. Reactome pathway analysis displayed the most enriched pathways. B-D HERC2-defcient Huh7 cells and HERC2-overexpressing HCC-97 h cells were treated with 50 ng/ml IL-6 (B), 50 ng/ml IL-11 (C), or 20 ng/ml EGF (D), respectively. The phosphorylation of JAK2 and STAT3 was determined by western blot analysis. E HERC2 and STAT3 double-defcient Huh7 cell lines were established. F The cells were treated with 50 ng/ml IL-6 for 24 h. The RT-qPCR assay was used to detect the mRNA expression of cancer stem cell-related genes. G The cells were cultured in a conditioned medium with 100×N2, 50×B27, 20 ng/ml EGF, 10 nmol FGF, 5 µg/ml insulin, and 0.4% BSA for 7 days, scale bars = 100 μm. H The cells were treated with 20 µM sorafenib for 24 h. A fow cytometry assay was used to determine the percentage of apoptotic cells. I Cells were treated with 50 ng/ml IL-6 for 24 h and then subjected to western blot assay for PD-L1 detection. J and K Activated PBMCs were cocultured with HCC cells at the ratio of 4:1 for 24 h. Apoptosis of HCC cells was detected by fow cytometry assay (J). K IFN-γ levels of CD8 + T cells were determined by fow cytometry analysis. NS: not signifcant, **p < 0.01, ***p < 0.001. Data from one representative experiment of three independent experiments are presented