Figure 2. Ketohexokinase knockdown improves metabolic dysfunction in HFD+F-fed mice.

A) mRNA expression of genes regulating first, as well as B) second and third steps of fructose metabolism and genes responsible for C) endogenous fructose production in the livers of male, C57BL/6J mice fed chow diet on regular water (Chow), or HFD supplemented with 30% fructose and injected with PBS (HFD+F/PBS) or injected with KHK siRNA (HFD+F/siKHK) for the last 8 out of 20 weeks on the diets. Chow group for every gene was normalized to 1. Hepatic D) triglycerides (TG) and total cholesterol levels in the livers of these mice. E) NAFLD activity score based on the interpretation of liver histology by an independent laboratory. Serum F) total cholesterol, as well as G) alanine aminotransferase (ALT) and aspartate aminotransferase (AST) obtained at sacrifice, n=8–11 mice per group. H) Glucose tolerance test in these mice performed after 18 weeks on the diet. Western blot quantification of liver KHK and proteins mediating I)de novo lipogenesis (ACLY, ACC1, FASN, SCD1) and J) ER stress (XBP1, PERK, CHOP) pathways. Sample number for WB is n=3–8 and for mRNA n=6 mice. Statistically significant results are marked with # using one-way ANOVA analysis, with Dunnett’s multiple comparisons test to the Chow group, # p<0.05; ## p<0.01; ### p<0.001; #### p<0.0001, while * represents a post-hoc analysis between the groups under the line, *p <0.05; ** p<0.01, *** p<0.001, **** p<0.0001. All data are presented as mean ± S.E.M.