Extended Data Fig.10. Requirement of g-beige fat for adaptive thermogenesis in the absence of β-AR signaling.

a, Schematic of the experiment. Pparg^MyoD KO mice (Myod-Cre^ERT2;Pparg^flox/flox) and littermate control mice (Pparg^flox/flox) were pretreated with β-blocker and tamoxifen. Subsequently, these mice were acclimated to 15°C for 5 days. b, Immuno-fluorescent staining of UCP1 and ENO1 in the inguinal WAT of Pparg^MyoD KO mice and controls. Scale bar=100 μm. The images represent three independent experiments. c, Quantification of glycolytic beige fat in (b). n=10. d, OCR in the inguinal WAT of Pparg^MyoD KO mice and littermate controls. n=6. e, EMG measurement of skeletal muscle shivering in Pparg^MyoD KO (n=6) and littermate control mice (n=5) at 30ºC or 15ºC. The shivering data were converted to RMS (μV). n.s., not significant. Data are mean ± SEM of biologically independent mice, and analyzed by ANOVA followed by Tukey’s test. f, Whole-body oxygen consumption (VO₂) in Pparg^MyoD KO mice and littermate controls. Mice were treated with β-blocker treatment at indicated time points at 15°C. n=5 for Pparg^MyoD KO mice, n=6 for control. Data are expressed as mean ± SEM of biologically independent mice, and analyzed by two-way ANOVA followed by Bonferroni’s test. g, Total food intake in (f). h, Locomotor activity in (f). i, Body weight of Pparg^MyoD KO mice and littermate controls on a regular chow diet. Mice were treated with β-blocker and acclimated to 15°C for 5 days. n=5 for Pparg^MyoD KO mice, n=7 for littermate control mice. (c,d,g-i) Data are mean ± SEM of biologically independent samples, and analyzed by unpaired two-sided Student’s t-test.