Fig 2: Inhibition of mitochondrial fission with Mdivi-1 attenuates LPS-induced STING signaling activation in KCs and protects liver function. Primary hepatocytes and KCs were isolated from WT mice treated with Mdivi-1 20 mg/kg or Vehicle (equal volume of DMSO) in the absence or presence of LPS (10 mg/kg for 12 h). (A–E) Western blot analysis of STING, IRF3, p-IRF3, p65, p-p65, DRP1, p-DRP1 protein expression in KCs (n = 6). (F–H) Levels of inflammatory cytokines including IFN-ß, TNF-a and IL-1ß in mouse serum in each group (n = 4). (I–J) Hepatocyte death detection by flow cytometry in each group (n = 4). (K–L) H&E staining and histology scores of liver sections in each group (n = 6, bar = 100 µm). (M - N) ALT and AST concentrations in mouse serum in each group (n = 6). Data are expressed as mean ± SEM, *p < 0.05, **p < 0.01, ***p < 0.001.

Fig 3: STING signaling pathway is activated in KCs in LPS-induced septic mice. (A–B) Representative images of immunohistochemistry (IHC) staining for STING protein in Control and LPS (10 mg/kg for 12 h) group. The AOD value of IHC staining was measured by imagePro Plus software (n = 6, 10 + fields per group, bar = 100 µm). (C) Western blot analysis of STING protein expression in KCs and hepatocytes isolated from WT mice (n = 6). (D–G) Western blot analysis of STING, IRF3, p-IRF3, p65 and p-p65 protein expression in KCs isolated from WT mice after Control or LPS treatment (n = 6). (H–J) The mRNA levels of inflammatory cytokines including IFN-ß, TNF-a and IL-1ß in KCs after Control or LPS treatment (n = 6). Data are expressed as mean ± SEM, *p < 0.05, **p < 0.01, ***p < 0.001.

Fig 4: TMEM173 (STING) and IRF3 are excessively expressed in liver fibrosis tissues from liver cancer patients.(A) The GSE number and sample quality for RNA microarray analysis of liver cancer tissue (tumor tissue, n=4) and normal adjacent tissue (ctrl, n=4) from GSE99807. (B) A volcano map derived from the differentially expressed genes (screening condition, p<0.05) between liver cancer tissue (tumor tissue, n=4) and normal adjacent tissue (ctrl, n=4) from GSE99807. (C) Using the differentially expressed genes between liver cancer tissue (tumor tissue, n=4) and normal adjacent tissue (ctrl, n=4) from GSE99807 to perform KEGG signal enrichment. (D) Using the differentially expressed genes in liver cancer tissue (tumor tissue, n=4) and normal adjacent tissue (ctrl, n=4) from GSE99807 to perform GOBP enrichment. (E and F) The expression distribution of TMEM173 and IRF3 genes in tissues (G1: normal, n=6; G2: liver fibrosis, n=6) from GSE33650, the upper left corner shows the significance p-value test method. COL1A1, collagen type I alpha 1 chain; STING, stimulator of interferon genes; IRF3, interferon regulatory factor 3.