a Graphic illustration of the experimental exercise training model. Mice were acclimatized to housing temperature before completing a 6 weeks voluntary wheel running exercise training (ET) intervention. (UT = untrained). b The effect of housing temperature and ET at 22 °C and 30 °C on body weight. Statistical testing; Two-way ANOVA (repeated measures). 22 °C UT; n = 8, 22 °C ET; n = 10, 30 °C UT; n = 9, 30 °C ET; n = 10. c–e VO2 and RER in UT and ET mice at 22 °C and 30 °C housing. Statistical testing; Two-way ANOVA (repeated measures). 22 °C UT; n = 5, 22 °C ET; n = 6, 30 °C UT; n = 8, 30 °C ET; n = 6. Effect of time within group; $$$p < 0.001. Effect of temperature within time of day; #p < 0.05, ##p < 0.01, ###p < 0.001. Effect of ET within temperature; **p < 0.01, ***p < 0.001. f Habitual activity (2 consecutive days) in UT mice after 6 weeks temperature acclimatization. Statistical testing; Two-way ANOVA (repeated measures). 22 °C UT; n = 5, 30 °C UT; n = 8. Effect of time within group; $$$p < 0.001. Effect of temperature within time of day; #p < 0.05. g Running distance per day at 22 °C and 30 °C, respectively. Statistical testing; Two-way ANOVA (repeated measures). n = 10. Effect of temperature; ###p < 0.001. h–k Effect of temperature on wheel running distance, maximum and average speed. Statistical testing; student’s t-test. 22 °C ET; n = 6, 30 °C ET; n = 6. Effect of temperature; #p < 0.05, ##p < 0.01. l Core temperature was measured at day (light period) and night (dark period) time via rectal thermometer. Statistical testing; Two-way ANOVA (repeated measures). n = 8 Effect of time, day vs. night; $p < 0.05, $$p < 0.01. Effect of temperature within day; #p < 0.05. Data are presented as mean ± SEM incl. individual values where applicable. The “n = x” defines the number of biologically independent animals used for the analyses.