Figure 1. Mice fed VHF/S developed obesity and insulin resistance.

In two separate studies, mice fed C (n = 14), VHF (n = 43) or VHF/S diet (n = 38) were monitored for 8 weeks and assayed for various metabolic parameters. (A) Body weight gain (14–43 mice per group). (B) Energy intake (14–43 mice per group). (C) Quantification of adipose tissue. Total fat pad includes epididymal, retroperitoneal and inguinal fat pad (7–16 mice per group). (D) H&E staining showing representative morphology of adipocyte in epididymal fat of animals (4–5 mice per group). (E) Glucose tolerance test. Glucose was injected and blood glucose was assessed at indicated time points (7–13 mice per group). (F) Glucose-stimulated insulin release. Plasma insulin levels were measured before and 15 min after injection of glucose in mice (8–12 mice per group). (G) Insulin tolerance test. Random-fed mice were injected with insulin and blood glucose assessed at indicated time points (7–13 mice per group). (H) Blood glucose (4–7 mice per group) (I) Plasma insulin (4–6 mice per group). (J) Muscle glucose uptake. Ex vivo soleus muscles were incubated without or with insulin and glucose uptake assessed (7–12 mice per group). (K) In vivo insulin signaling. Overnight fasted animals (n = 4–5 per group) were injected with insulin or saline and expression of Akt and pAkt in gastrocnemius muscles was assessed. Graphic depicts densitometric analysis of normalization of pAkt/Akt protein. Representative western blots of muscle lysates are shown for phosphorylated Akt (Ser⁴⁷³) without or with insulin stimulation, and for total Akt expression after saline injection. Western blot analyses were repeated at least three times. (L) Triacylglyceride (TAG) concentrations in gastrocnemius muscles (6–12 mice per group). *p<0.05 vs. C. **p<0.03 vs. VHF.