Thermoneutrality reverses the protective phenotype observed in miPEP deficient mice. (A) Raw energy expenditure (Kcal/h) for adipo-miPEP-KO (aKO, light blue) and miPEPfl/fl (black) mice housed at room temperature over 48 h period (B) Quantification of energy expenditure ANCOVA adjusted during light and dark circles every 24 h Mean ± S.D., N = 4. p-value reported (C) Change in adiposity in miPEPfl/fl (fl/fl, grey) and adipo-miPEP-KO mice (aKO, light green) in response to a high fat, high sugar diet (HFHSD) in thermoneutrality (TN) or room temperature (RT) relative to baseline. Mean ± S.D., N = 4–7, ∗p < 0.05, ∗∗∗p < 0.001 vs miPEPfl/fl mice. (D) Oral Glucose tolerance test, (E) Blood insulin in fasting and at 15 min during the oral glucose tolerance test and (F) HOMA-IR after 4 wks of HFHSD in thermoneutrality (TN) or age-matched mice housed at room temperature (RT). Mean ± S.D., N = 4–7. ∗p < 0.05. Mean ± S.D., N = 4. p-value reported. (G) Mice were housed at thermoneutrality (TN) and fed a HFHSD for 4 weeks. Following this period, their EE was measured over 48 h under TN. EE was quantified (ANCOVA adjusted). Data are presented as mean ± S.D. (N = 4). The p-value is reported.