Fig 2: SNS-ameliorated LPS-induced inflammatory injuries and cytotoxicity. Mice were injected with 0.25 mg/kg/day LPS for 4 days, or 0.25 mg/kg/day LPS and SNS (low dose at 12.5 mg/kg/day, medium dose at 25 mg/kg/day, high dose at 50 mg/kg/day) for 4 days. (A) Changing of the ratio of liver/body weight (%) after LPS with/without SNS treatments. (B) Serum ALT and AST levels after indicated treatment. (C) H&E staining and TUNEL staining in the liver and lungs after indicated treatment. n = 6, *p < 0.05, ***p < 0.001, ****p < 0.0001.

Fig 3: Testosterone alleviates histopathological and biochemical findings of liver fibrosis. Liver injury was induced in C57/BL male mice for 2 and 4 weeks and was compared with naive counterparts. Testosterone was administered via i.p injection for 1 week and 2 weeks, starting at weeks 1 and 2 of the acute and chronic CCl4 models, respectively, as described in Materials and Methods. Representative images of immunohistochemical liver staining sections of (A) H&E and (B) Sirius red, shown at an original magnification of ×10. The quantification of liver histology assessments is presented in (C) as the average ±SD for each group (six mice per group). Serum markers of liver injury (D) ALT and (E) AST were measured. mRNA markers of liver fibrosis of (F) αSMA and (G) collagen III were assessed. Each experiment was repeated three times to ensure reliability and reproducibility. [**p = 0.01, ***p = 0.005, and ****p = 0.0001]. A Mann-Whitney U test was performed to evaluate whether the liver injury mice marker (ALT and AST) was altered by testosterone treatment in both the acute and chronic CCl4-injected groups. The results demonstrated significant values in both groups. Therefore, the null hypothesis is rejected. Data is normally distributed (alpha = 0.05).

Fig 4: Combined targeting both ITGB1 and ITGB4 inhibits pUSP14‐MEF2D‐ITGB1/4 signaling and intrahepatic metastasis of disseminated HCC cells. A,B) IHC staining (A) of MEF2D, ITGB1, ITGB4, pY397‐FAK or pS432‐USP14 and quantitative analysis of their staining scores (B) in HCC tissues from 75 patients (Cohort II). Representative IHC images from two samples are shown. Scale bar: 50 µm. C) HCC patients were stratified for Kaplan‐Meier analysis of overall survival (OS), relapse‐free survival (RFS) and progression‐free survival (PFS) according to the co‐expression levels of MEF2D, ITGB1 and ITGB4 in tumour tissues using data from the TCGA dataset. D) Schematic of anti‐ITGB1, anti‐ITGB4, or combination treatment strategy of mice implanted with HCCLM3 cells through splenic or intrahepatic injection (n = 6). E) BLI at 1, 21, and 60 days post splenic injection of HCCLM3 cells. F) Livers resected from the spleen‐to‐liver metastasis mouse model. Tissues were photographed, fixed, and stained with haematoxylin and eosin (H&E), or immunostained for the levels of MEF2D, ITGB1, ITGB4, pY397‐FAK, and pS432‐USP14 as indicated (n = 6). Black arrow, liver metastases. Scale bar, 1 cm (top), 100 µm (bottom). G‐H) Liver metastases were counted (G) and plasma ALT and AST levels were measured (H) for mice. For all panels, * p < 0.05, ** p < 0.01, *** p < 0.001, and ns, no significance.

Fig 5: Antitumor activity of rencofilstat in HCV-induced HCC: (A) Humanized MUP-uPA-SCID/Beige mice (n = 10) were infected with HCV GT1a and daily treated with rencofilstat starting at week 0, 12, 16, and 24 post-infection. Liver nodules were analyzed at week 24 post-HCV infection. Liver nodules were counted, their diameters were measured and the cancerous status of each nodule was verified via qRT-PCR for tumor markers MCP-1, Timp-1, and glypican-3. (B) Statistical analysis was conducted. (C) HCV GT1 infection was monitored via qRT-PCR at the indicated time points. Data are expressed as HCV RNA copies/mL of serum. (D) Each mouse from each group was weighed every 4 days. (E,F) AST and ALT serum levels (units/liter) were quantified with ELISA at the indicated time points.