Prebiotic consumption reduces hepatic steatosis and cholesterol levels independent of body weight regulation. (A) Outline of the three different prebiotic feeding regimes used in this study. The three groups have different access to prebiotics; HF = no access to prebiotics, HF-UP = unrestricted access to prebiotics and HF-ARP = active phase (ZT12-ZT0) restricted access to prebiotics. During the experimental period all three groups had ad libitum access to HFD. (B) Daily food intake during the study period. (C) Prebiotic intake during the study period. (D) Body weight gain over time. (E) Final body weight gain (n = 10). (F) Quantification of serum cholesterol (n = 8/group). (G) Representative histology images of H&E-stained liver sections. (H) Quantification of hepatic triglyceride (n = 10/group except HF-ARP n = 9). (I) Relative mass of liver (n = 10). Data expressed as mean ± SEM and statistical significance assessed by two-way repeated measures ANOVA (B, D) ANOVA one-way ANOVA (E-F, H-I), followed by a Tukey post-hoc test for multiple group comparison and unpaired t test for (C) (*p < 0.05, **p < 0.01, ***p < 0.001).