Figure 1.

Treatment of hu-liver NASH mice with GalNAc-siTAZ lowers liver inflammation and fibrosis

(A) Experimental scheme. Humanized mice were fed the HF-CDAA diet for 6 weeks and then injected once weekly for 6 additional weeks with 5 mg/kg GalNAc-siTAZ (siTAZ) or control GalNAc construct (control) while still on the diet. At 12 weeks, the 2 cohorts were analyzed for systemic and liver endpoints. (B) Body weight. (C) Livers were immunostained with anti-STEM121, and the percentages of hepatocytes that were STEM121⁺ were quantified; liver from a mouse fed the HF-CDAA diet for 12 weeks was used as a negative control for anti-STEM121 staining. Scale bar, 100 μm. (D) Liver WWTR1 and IHH mRNA and immunoblots of YAP, TAZ, IHH, and β-actin; a longer exposure (long) of the TAZ immunoblot is included. The densitometric data for IHH is shown. (E) Livers were assayed for the inflammatory mRNAs Cxcl9, Tnfa, Emr1, Tgfb1, and Mcp1 and the fibrosis-related mRNAs Spp1, Acat2, and Col1a1 mRNAs. (F) Livers were stained with H&E (upper row) and Sirius red (lower row) and then quantified for inflammatory cells per field and percentage of Sirius red area. Scale bar, 200 μm. (G) Livers were immunostained with anti-αSMA and quantified for the αSMA⁺ area, which was normalized for cellularity by dividing by the number of DAPI⁺ cells. Scale bar, 200 μm. (H) Plasma ALT and AST. (I) Percentage of lipid droplet area in the livers. (J) Liver NOTCH1 and HES1 mRNA. Values shown for all of the graphs are means ± SEM; n = 6 control mice and 7 siTAZ mice. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001. The data were analyzed using the Student’s t test.