Table 1.
Sequences of primers and probes used for real-time PCR.
Figure 1.
Effects of cGFJ/sweetener on liquid consumption.
Mice were given 100% cGFJ, 100% cGFJ +0.15% saccharin, 50% cGFJ +0.15% saccharin +1.5% cyclamate or 50% cGFJ +0.15% saccharin as their sole liquids: A) liquid consumption, ***P = 0.0007; B) liquid consumption, ***P<0.0001; C) liquid consumption, *P<0.02; D) liquid consumption, P>0.6. Statistical significance was determined by two-tailed, unpaired t-tests.
Figure 2.
Effects of cGFJ on liquid and food intake, and weight.
Mice were fed a LFD or HFD and 50% cGFJ for 100 days, starting from weaning (day 0) at 4 wk old. LFD: A) cumulative liquid consumption; B) cumulative food consumption; C) total body weights; D) intra-abdominal fat pad weight. HFD: E) cumulative liquid consumption; F) cumulative food consumption, *P<0.05; G) total body weights, ***P = 0.0001; H) intra-abdominal fat pad weight, P<0.05; I) caloric content of feces of cGFJ and control water treated mice fed a HFD for 100 days, P>0.7 and fecal weight of mice collected over 24 hr at the end of 106 days of treatment, P>0.03 for GFJ. A two-tailed, unpaired t-test was used to determine statistical significance.
Figure 3.
cGFJ effects on blood glucose and insulin sensitivity.
Mice were treated as described in the legend of Fig. 2. A, B) values at the end of 100 d LFD: A, fasting blood glucose; B) fasting serum insulin, *P<0.04; C, D) GTT and AUC of mice fed a HFD at week 13, *P<0.04; E, F) ITT and AUC of mice fed a HFD at week 11, ***P<0.001. G, H) Values at the end of 100 d HFD: G) fasting blood glucose, *P<0.02; H) fasting serum insulin, *P<0.03. A two-tailed, unpaired t-test was used to determine statistical significance.
Figure 4.
Impact of cGFJ on AKT activity in liver and skeletal muscle, TG content, and fatty acid synthesis.
Mice were fed a HFD and 50% cGFJ for 100 d as described in the legend of Fig. 2. A) pAKT/total AKT ratios in muscle (***P = 0.0002) and liver (*P<0.05); B) liver TG, *P<0.05; I and II) Representative sections from control water and GFJ treated animals, respectively; C) total FFA synthesis and individual FFA; D) GTT and AUC at week 6, P<0.005. A two tailed, unpaired t-test was used to determine statistical significance.
Figure 5.
Impact of cGFJ on established diet-induced obesity.
Mice were fed a HFD for 6 wk starting at 4 wk old. Animals were then divided randomly into control and GFJ groups (day 0) and HFD feeding was continued an additional 56 d: A) cumulative liquid consumption; B) cumulative food consumption; C) total body weights; D) blood glucose; E) GTT and AUC at week 6, P<0.002; F) ITT and AUC at week 7, P<0.03. A two tailed, unpaired t-test was used to determine statistical significance.
Figure 6.
Comparison of metabolic effects of cGFJ, naringin, and metformin.
Mice fed a HFD were given cGFJ, water containing naringin or metformin, or control water for 106 d: A) cumulative liquid consumption and rates based on linear regression. Slopes did not differ significantly; B) cumulative food consumption and rates based on linear regression. Slopes did not differ significantly; C) total body weights. 2way ANOVA with Bonferroni posttests showed that treatment and time had a significant effect (p<0.0001) on body weight. Bonferroni posttests only showed significant differences in body weight between the control and cGFJ group form 92 onward but not for any of the other groups; D) blood glucose. 2way ANOVA with Bonferroni posttests showed that treatment and time had a significant effect (p<0.0001) on blood glucose levels. Bonferroni posttests comparisons to the water control group showed significant differences for cGFJ and naringin starting day 27 and metformin starting day 8. Differences between cGFJ, naringin, and metformin were non-significant at all time points.
Figure 7.
Effects of cGFJ on serum glucose levels and on AMPK and ACC phosphorylation in muscle and liver.
Mice were fed a HFD. A) Blood glucose levels during 17 d access to 50% cGFJ, 25% cGFJ, metformin, metformin and 50% cGFJ, or water. B-E) Mice were allowed access to 50% GFJ, naringin or metformin in control water for 106 d starting at weaning: B) liver pAMPK/AMPK, *P<0.04, ***P = 0.0002; C) liver pACC/ACC, *P<0.03; D) muscle pAMPK/AMPK, *P<0.03; E) muscle pACC/ACC, P<0.03. Phospho- and total protein were determined by western blot and normalized to tubulin. A two-tailed, unpaired t-test was used to determine statistical significance.
Figure 8.
Gene expression changes caused by cGFJ.
Mice were allowed to drink 50% cGFJ or control water for 17 or 100 d, as described in the legend of Fig. 2. A) liver, short term; B) small intestine, short term; C) liver, long term; D) small intestine, long term. Values were normalized to control values set as 1: *P<0.05.