(A–C) Energy expenditure, (D–F) cumulative food intake and (G–I) respiratory exchange ratio (RER) in 10–14 week old, ad libitum fed, wild-type and ACC DKI male mice after 72 hr of exposure to indicated temperatures (n = 8). (A, B, D, E, G, H) Hourly averages of metabolic parameters of mice exposed to 14°C and 28°C. Black horizontal bars represent the dark period in a 12 hr light/dark cycle (7am-7pm). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 represent differences between genotypes for a given time point as determined by 2-way repeated measures ANOVA with Bonferroni post-hoc test. (C, F, I) 12 hr averages of at least two consecutive day and night cycles of metabolic parameters of mice exposed to indicated temperatures. *p<0.05, ****p<0.0001 differences between genotypes within a temperature group and; # p<0.05, p<0.0001 differences between parameters for a given genotype at different temperatures as determined by 2-way ANOVA with Bonferroni post-hoc test. (J) mRNA expression profile in brown fat of mice exposed to 14°C for 72 hr (n = 12). *p<0.05, **p<0.01 genotype difference in mRNA expression for a given gene as determined by unpaired t-test, two-tailed. (K) Percentage body weight change of wild-type and ACC DKI mice exposed to various temperatures for 72 hr. *p<0.05 represents differences between genotypes and p<0.001, p<0.0001 represent differences in body weight loss at different temperatures as determined by 2-way ANOVA and Bonferroni post-hoc test. All data are presented as mean ± s.e.m.