Figure 2.

A repeat experiment showing that GalNAc-siTAZ lowers NASH progression in hu-liver mice

A second cohort of hu-liver mice were treated with GalNAc-siTAZ or control GalNAc construct exactly as depicted in Figure 1A. (A) 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. (B) Liver WWTR1 and IHH mRNA and immunoblots of TAZ, IHH, and β-actin. (C) Livers were immunostained with anti-TAZ and anti-STEM121 and then quantified for the percentage of TAZ⁺cells among STEM121⁺cells. Scale bar, 200 μm. (D) 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. (E) Livers were immunostained with anti-αSMA and quantified as in Figure 1G. Scale bar, 200 μm. (F) Hydroxyproline content. (G) Livers were assayed for the inflammatory mRNAs Emr1, Mcp1, Tnfa, Cxcl9, and Tgfb1 and the fibrosis-related mRNAs Timp1, Col1a1, Col3a1, Col1a2, and Acta2. (H) Livers were immunostained with anti-Mac2 and anti-Clec4f and quantified for the percentage positive area of each. Scale bar, 200 μm. (I) Livers were immunostained with anti-F4/80 and quantified for the percentage of positive area. Scale bar, 200 μm. (J) Plasma ALT. (K) Percentage of lipid droplet area in the livers. (L) Livers were immunostained with anti-Ki67 and quantified for the percentage of positive area. Scale bar, 200 μm. (M) Body weight. (N) The livers of a subgroup mice from each cohort (5 control, 4 siTAZ) with similar body weights were assayed for percentage of Sirius red⁺. (O) Liver NOTCH1, HES1, and EPHB2 mRNA. Values shown for all of the graphs are means ± SEM; n = 8 mice per group. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001. The data were analyzed using the Student’s t test.