Effect of MHB on plasma components in mice fed on HFD with or without MHB supplementation.

<p>Data are presented as means ± SEM. n = 12 mice/group.</p><p>**<i>P</i> < 0.01 (by unpaired Student’s <i>t</i>-test). NEFA, non-esterified fatty acid.</p><p>Effect of MHB on plasma components in mice fed on HFD with or without MHB supplementation.</p

Total food intake of mice fed on HFD with or without MHB supplementation and the effect of MHB on fecal lipid content.

<p>Data are presented as means ± SEM. n = 12 mice/group. No significant differences were observed by unpaired Student’s <i>t</i>-test.</p><p>Total food intake of mice fed on HFD with or without MHB supplementation and the effect of MHB on fecal lipid content.</p

Single oral administration of MHB elevated BAT-SNA in rats.

<p>(A) Time course of BAT-SNA changes taken every 5 min and mean BAT-SNA over 0- to 90- min period. Urethane-anesthetized rats were administered 10 mg/kg of MHB. Data were calculated as a percentage of baseline, and as means ± SEM. n = 3 rats/group. **<i>P</i> < 0.01 (by ANOVA with repeated measures). (B) Effects of 2 mg/kg MHB on mean BAT-SNA over 0- to 90- min period in sham-operated or vagotomized rats. Data are presented as means ± SEM. n = 3 rats/group. **<i>P</i> < 0.01 (by ANOVA with repeated measures).</p

Measurement of mRNA expression levels and Immuno-blot analysis in the interscapular BAT of mice fed HFD supplemented with MHB.

<p>(A) mRNA expression levels in BAT. mRNA expression levels were normalized to the expression level of <i>GAPDH</i> as a reference. (B) Immuno-blot analysis of UCP1 in BAT mitochondria. Representative immuno-blots are shown on the histogram. Representative signals shown are from the same immuno-blot membrane. UCP1 protein level was normalized to the level of UQCRC1 protein as a reference. Data are presented as means ± SEM. n = 12 mice/group. *<i>P</i> < 0.05, **<i>P</i> < 0.01 (by unpaired Student’s <i>t</i>-test).</p

MHB ameliorated HFD-induced body fat accumulation in mice.

<p>(A) Body weight gain of HFD-fed mice with or without MHB supplementation. Data are presented as means ± SEM. n = 12 mice/group. **<i>P</i> < 0.01 (by analysis of variance (ANOVA) with repeated measures). (B) Epididymal WAT, abdominal subcutaneous WAT and liver weight. Data are presented as means ± SEM. n = 12 mice/group. **<i>P</i> < 0.01 (by unpaired Student’s <i>t</i>-test).</p

Single oral administration of MHB increased the level of cAMP in BAT and elevated its temperature in rats.

<p>(A) cAMP level in the lysates of BAT obtained from rats administered MHB pretreated with propranolol or saline. Thirty minutes after the pretreatment with 10 mg/kg propranolol or saline, rats were administered 10 mg/kg MHB and killed after 3 h. Data are presented as means ± SEM. n = 10 rats/group. **<i>P</i> < 0.01, N.S., not significant (by unpaired Student’s <i>t</i>-test). (B) Time course of BAT temperature changes (ΔT_BAT) taken every 1 min relative to the baseline. The baseline was determined as the mean T_BAT over a 5-min period before the administration of MHB. Urethane-anesthetized rats were administered 10 mg/kg of MHB. Data are presented as means ± SEM. n = 4 rats/group. **<i>P</i> < 0.01 (by ANOVA with repeated measures).</p