Extended Data Fig. 4: Maintenance of adiposity in female mice under cold exposure.

a) Images showing cell morphology of beige differentiated SVFs isolated from male and female iWATs. Scale bar at 100 μm. b) Real-time qPCR of indicated genes from the scWATs of 24 h cold exposed male and female adrenalectomized mice. N = 6,6. c–e) Individual mice data for oxygen consumption (VO2 ml/hr), and energy expenditure (EE kcal/hr) (c), respiratory exchange ratio (RER) and generalized linear model (GLM)-based regression plots of RER with total body weight (Total), lean mass (Lean) and fat mass (Fat) as co-variates (Two-way ANCOVA) showing p-value for Mass effect, Group effect, and Interaction effect (d), and total food consumed (kcal), locomotor activity (beam breaks), and total distance in cage (m) (e) of male and female mice exposed to 22 °C for 21 h and 4 °C for 24 h in Sable Promethion metabolic chambers (12 h light/dark cycle, 45 h total duration, white bar represents light cycle and grey bar represents night cycle). N = 6,6. f and g) Body weight (f) and lean mass (g) of male and female mice before (RT) and after 24 h cold exposure (COLD). N = 6,6. h) Weights of indicated fat depots in cold exposed male and female mice. N = 8,8. i and j) Gross appearance, H&E staining, and immunostaining of UCP1 of male dorsolumbar iWATs and female dorsolumbar mgWATs exposed to 24 h cold (i) and Western blot of UCP1 in iWAT from 24 h cold-exposed males and females (j). Actin was used as a loading control and UCP1 bands were quantified and normalized to Actin intensity. N = 4,4. Scale bar at 200 μm. Unpaired Student’s t Test (b,f,h) and ANCOVA (d) *, p < 0.05, **, p < 0.01; ***, p < 0.001, n.s., not significant. Data are represented as mean ± S.E.M.

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