Table 1.
Primer sequences for RT-qPCR
Fig 1.
Characterisation of the effect of diet composition on C57BL/6 mice.
(A) Effect of Chow, HCD and HFD on body weight over 12 weeks (n = 5 per group). (B) Effect of Chow, HCD and HFD on percentage body fat over 12 weeks, as assessed by DEXA scan (n = 5 per group). (C) RQ and (D) Energy expenditure from a representative day and night. Data from CLAMS metabolic cages after 12 weeks on diet (n = 4 per group). (E) Food intake and (F) Energy intake averaged over 12 weeks (n = 5 per group). Results are expressed as mean ± SEM. Statistical analysis was by two-way repeated measures ANOVA followed by Dunnett’s post-hoc test (A and B) or a one-way ANOVA followed by Dunnett’s post-hoc test (C, D and E). (*: p<0.05, ***: p<0.001) (a: p<0.05 vs Chow group, b: p<0.05 vs HCD group)
Fig 2.
A high-calorie, high-carbohydrate diet results in glucose intolerance but improved insulin sensitivity.
(A) Glucose tolerance test (GTT), (B) Area under the curve (AUC) of GTT, (C) Insulin tolerance test (ITT), and (D) AUC of ITT. Tests performed after 12–14 weeks on diet. (E) GTT and (F) ITT percentage change after 2 weeks on diet. (G) Serum insulin. Statistical analysis was by two-way repeated measures ANOVA followed by Dunnett’s post-hoc test (A, C, E and F), one-way ANOVA followed by Dunnett’s post-hoc test (B, D), or two-way ANOVA followed by Dunnett’s post-hoc test (G). (**: p<0.01, ***: p<0.001, ****: p<0.0001) (a: p<0.05 vs Chow group, b: p<0.05 vs HCD group, c: p<0.05 HCD vs chow, d: p<0.05 HFD vs chow)
Fig 3.
Effect of diet on sensitivity to exogenous insulin in C57BL/6 mice.
(A) Serum non-esteried fatty acid (NEFA) levels 10 minutes after a saline or insulin injection (5U/mouse) (n = 3–4 per group). (B) Pancreatic insulin content after 12 weeks on diet (n = 13–14). Western blotting for Akt, p-Ser473-Akt in liver (C), WAT (E) and gastrocnemius muscle (G) of mice after a 4 hour fast followed by either a saline or insulin injection (5U/mouse) 10 minutes before sacrifice. Band density was analysed using Image J and pAkt/Akt ratio was calculated and graphed using chow/saline as control (D, F, H). Results are expressed as mean ± SEM. Statistical analysis was by two-way ANOVA followed by Dunnett’s post-hoc test (A, D, F, H). (*: p<0.05, **: p<0.01 vs sham controls) (a: p<0.01 vs sham controls, b: p<0.01 vs HFD insulin)
Fig 4.
A high-calorie, high-carbohydrate diet results in liver steatosis.
(A) A representative photo of a liver from mice fed a chow or a HCD. (B) Effect of diet on liver triglyceride levels in mice. (C) Representative Oil Red O and H+E liver sections from mice fed either a chow, HCD or HFD for 12 weeks. Magnification = 20x. Results are expressed as mean ± SEM. Statistical analysis was by one-way ANOVA followed by Dunnett’s post-hoc test (B). (**: p<0.01 vs chow mice)
Fig 5.
Effect of diet on ChREBP gene expression and the lipogenic pathway.
Effect of diet on gene expression in liver (A), skeletal muscle (B), white adipose tissue (C) and ileum (D). N = 5 per group. Results were normalized to expression of the housekeeping gene 36B4, and then shown as fold change versus the chow control mice. Results are expressed as mean ± SEM. Statistical analysis was by two-way ANOVA followed by Dunnett’s post-hoc test. (*: p<0.05, **: p<0.01, ***: p<0.001, ****: p<0.0001 versus chow fed mice)
Fig 6.
Effect of diet on inflammatory gene expression and serum inflammatory markers.
Effect of diet on gene expression in liver (A), WAT (B) and ileum (C) and serum resistin (D) and PAI-1 (E). N = 5 per group. For gene expression, results were normalized to expression of the housekeeping gene 36B4, and then shown as fold change versus the chow control mice. Results are expressed as mean ± SEM. Statistical analysis was by two-way ANOVA (A, B, C) or one-way ANOVA (D, E) followed by Dunnett’s post-hoc test. (*: p<0.05, **: p<0.01, ***: p<0.001, ****: p<0.0001 versus chow fed mice)