Fig 2: Treatment of high-fat-diet–fed mice with GalNAc-Stk25 ASO markedly reduced hepatic STK25 levels. Mice were treated with GalNAc-Stk25 ASO (12.5 mg/kg/wk), Stk25 ASO (50 mg/kg/wk), GalNAc-control ASO (12.5 mg/kg/wk), or placebo (PBS) for 6 weeks. (A) Hepatic Stk25 mRNA expression. (B–D) STK25 protein abundance in the (B) liver, (C) skeletal muscle, and (D) WAT. Protein levels were analyzed by densitometry; representative Western blots are shown with pan-actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), or ß-actin used as loading controls. (E) Representative liver sections processed for immunofluorescence with anti-STK25 antibody (red); nuclei were stained with DAPI (blue). Scale bars: 20 µm. (A–D) Data are means ± SD (n = 6–8 mice per group) compared with a control group of mice dosed with PBS. Cntr, control. *P < .05, **P < .01.

Fig 3: Measurement of mRNA expression of selected genes related to (A) hepatic inflammation and (B) fibrosis, as well as genes controlling (C) lipid synthesis and (D) oxidation in the livers from high-fat–fed mice. Mice were treated with GalNAc-Stk25 ASO (12.5 mg/kg/wk), Stk25 ASO (50 mg/kg/wk), GalNAc-control ASO (12.5 mg/kg/wk), or placebo (PBS) for 6 weeks. Relative mRNA expression was assessed by quantitative real-time polymerase chain reaction. Data are means ± SD (n = 6–8 mice per group) compared with a control group of mice dosed with PBS. Cntr, control. *P < .05, **P < .01.

Fig 4: STK25 disruption in mice protects against hepatic oxidative and ER stress in HCC. (A) Representative liver sections stained with dihydroethidium (DHE) (pink) or MitoTracker Red (red), or processed for immunofluorescence with anti–4-hydroxynonenal (anti–4-HNE) (green), anti-E06 (green), anti-KDEL (green), or anti–C/EBP homologous protein (anti-CHOP) (red) antibodies; nuclei stained with DAPI (blue). (B) Quantification of the staining. Scale bars: 50 µm; except for MitoTracker Red staining: scale bars: 25 µm. Data are means ± SEM from 6 to 10 mice per group. CD, chow diet; DAPI, 4',6-diamidino-2-phenylindole; HFD, high-fat diet; KO, knockout; WT, wild-type. **P < .01, ***P < .001.

Fig 5: Knockout of STK25 reduces tumor angiogenesis in mouse models of DEN- or CDAA-induced HCC. (A) Representative liver sections stained with H&E or processed for immunofluorescence with anti-CD31 (red) antibody; nuclei stained with DAPI (blue) in immunofluorescence images. (B) Staining with anti-CD31 antibody was selectively quantified in tumor tissue identified from the H&E staining on a serial section (shown by dashed lines). Scale bars: 50 µm. Data are means ± SEM from 7 mice per group. DAPI, 4',6-diamidino-2-phenylindole; HFD, high-fat diet; KO, knockout; WT, wild-type. ***P < .001.