Figure legends
Figure 1. (a) Study design. (b) Body weight development over the intervention period. (c) Food intake over the course of the study. ***p<0.001 vs DIO-NASH vehicle control.
Figure 2. Systemic safety and metabolic parameters. (a) Plasma ALT, (b) plasma AST activities. (c) Plasma total cholesterol, (d) plasma triglycerides, (e) 4-hour fasting blood glucose and (f) plasma insulin levels. (a) -(d) were measured at study termination, (e) and (f) on study day 39. *p<0.05, **p<0.01, ***p<0.001 vs DIO-NASH vehicle control.
Figure 3. Liver weight, hepatic lipid, collagen and α-smooth muscle actin (α-SMA) content at study termination. (a) Wet liver weight, (b) hepatic triglycerides, (c) hepatic total cholesterol. (d) Total liver lipid content, (e) hepatic collagen 1a1 and (f) liver α-SMA determined by histology and quantitative image analysis. **p<0.01, ***p<0.001 vs. DIO-NASH vehicle control.
Figure 4. (a) Representative images of liver morphology at termination of the study. H&E staining (20x, scale bar = 100 µm), (b) Picrosirius Red staining (20x, scale bar = 100 µm). (c) Change in NAFLD activity score (pre- vs. post-treatment) for individual animals in the different treatment groups: Lean control mice (grey), DIO-NASH vehicle controls (black) and DIO-NASH mice treated with resmetirom (orange), (d) Change in fibrosis stage as above. N=10-11 per group, **p<0.01 vs DIO-NASH vehicle group (One-sided Fisher’s exact test with Bonferroni correction).
Figure 5. Hepatic gene expression assessed by RNA sequencing with subsequent Ingenuity pathway analysis: (a) principal component analysis over all 53431 measured transcripts displaying the first two main components, PC1 and PC2. (b) Visualization of upstream factor analysis performed with Ingenuity, which predicts THR-ß as one important factor activated in the resmetirom treated group. (c-e) Expression of selected genes in normalized counts (FKPM) and shown as box-whisker blots with median (line) and mean (+) and confidence intervals of 5% to 95%. *p<0.05, **p<0.01, *p<0.001 vs DIO-NASH vehicle group (false-discovery rate adjusted for multiplicity by Benjamini-Hochberg correction).