Fig. 8
HERC2 promoted hepatic STAT3 activation and facilitated HCC tumorigenesis in vivo. A Diagram for mouse primary HCC induction. 15-day-old male mice were intraperitoneally injected with 25 μg/g DEN. Two weeks later, the mice were intraperitoneally injected with 0.5 μl/g CCl4 once a week for consecutive 22 weeks. B Tumor formation in liver tissues was observed, and the red arrow indicates typical tumor nodes. C The statistical diagram for tumor numbers is presented. D The statistical diagram for the liver index is presented. (E) H&E staining was performed to examine the pathological changes in liver tissues. Top scale bars = 250 μm, and bottom scale bars = 25 μm. F The expression of Ki67 and PCNA in liver tissues was determined by immunohistochemical assay. Scale bars = 100 μm. G Western blot analysis was performed to detect JAK2/STAT3 signaling activation. H RT-qPCR analysis was conducted to evaluate the levels of cancer stem cell-related genes. I Western blot analysis was performed to detect PD-L1 expression in liver tissues. J PD-L1 expression was evaluated by immunohistochemical assay. Top scale bars = 250 μm, and bottom scale bars = 50 μm. K The expression of CD8 was determined by immunohistochemical assay. Top scale bars = 250 μm, and bottom scale bars = 50 μm. L-N A total of 2 × 106 or 2 × 105 HERC2-overexpressing Hepa1-6 cells or control cells were orthotopically injected into mouse livers. L A diagram of mouse orthotopically implanted HCC is presented. M Tumor formation in liver tissues is presented. N A statistical diagram for liver tumor volume is presented. O-Q A total of 2 × 106 HERC2-overexpressing Hepa1-6 cells or control cells were orthotopically injected into mouse livers. O H&E staining was performed to examine the pathological changes in liver tissues. Top scale bars = 250 μm, and bottom scale bars = 25 μm. P The expression of Ki67 and PCNA in liver tissues was determined by immunohistochemical assay. Scale bars = 250 μm. Q CD133 expression in liver tissues was determined by immunohistochemical assay. Top scale bars = 100 μm, and bottom scale bars = 25 μm. *p < 0.05, **p < 0.01. Data from one representative experiment of three independent experiments are presented