Extended Data Fig.2. Myod-derived beige adipocytes in adipose tissue.

a, Immuno-fluorescent staining of GFP and UCP1 in the anterior, middle, and posterior regions of inguinal WAT of Myod-Cre^ERT2 reporter mice. Mice were pre-treated with β-blocker and acclimated to 15°C. Scale bar=100 μm. Note that GFP⁺ adipocytes were enriched in the middle part of inguinal WAT near the lymph node (LN). The ratio (%) of UCP1⁺ cells among total GFP⁺ cells are shown on the top. N.D., not detected. b, Immuno-fluorescent staining of GFP and CD31 in the middle part of inguinal WAT. Myod-Cre^ERT2 reporter mice treated with β-blocker. Scale bar=100 μm. Note that GFP⁺ adipocytes are localized near the CD31⁺ microvasculature. c, Immuno-fluorescent staining of GFP and UCP1 in the iBAT of Myod-Cre^ERT2 reporter mice treated with β-blocker or vehicle. Scale bar=100 μm. d, Immuno-fluorescent staining of GFP and UCP1 in the epididymal WAT of mice in (c). Scale bar=100 μm. e, Immuno-fluorescent staining of GFP and UCP1 in the inguinal WAT of Myod-Cre^ERT2 reporter mice that were acclimated to 6ºC for 2 weeks without β-blocker treatment. Scale bar=100 μm. f, Immuno-fluorescent staining of GFP and UCP1 in the inguinal WAT of Myod-Cre^ERT2 reporter mice treated with CL316,243 (1mg kg⁻¹ day⁻¹) for 2 weeks. Scale bar=100 μm. g, Quantification of GFP⁺ beige adipocytes among total UCP1⁺ beige adipocytes in (e) and (f). N.D., not detected. n=8. Data are expressed as mean ± SEM of biologically independent experiments. h, Schematic of the experiment. Myog-Cre;Rosa26-mTmG reporter mice were treated with β-blocker at room temperature for 5 days and subsequently acclimated at 15° C for 5 days. i, Immuno-fluorescent staining of GFP and UCP1 in the inguinal WAT of mice in (h). Scale bar=100 μm. (a-f,i) tdTomato or DAPI for counter-stain. The images represent three independent replicates.