Metabolic analysis
HFD-mice were fed either vehicle or HSG4112 (0.5% feed mixture) for 4
weeks before the metabolic rate analysis and additional 2 weeks for
confirmation of full weight reduction in 6-week timeframe; weight-loss
effect equivalent to 100
mg·kg-1·day-1 administration was
observed (Supplementary Figure 2 ). Starting from the evening of
the day before the metabolic analysis, no feed and only water was
provided. The mice were placed inside Oxymax/CLAMS (Columbus Inc., USA)
for 24 hours before the experiment for environmental adaptation and
maintenance of a comfortable state, and for additional 49 hours for the
experiment. The O2 and CO2 generated by
the mice’s breathing was measured along with the O2 and
CO2 of the air flowing in from the room and into the
device. All parameters – VO2, VCO2,
respiratory exchange ratio, energy expenditure, and locomotor activity
– were calculated using built-in software and previously reported
method (Nie, Gavin and Kuang, 2015).