Figure 8. SMA+ mural cells are required for beige adipocyte formation.

(a–d) Two-month-old SMA-Cre^ERT2; PPARγ^fl/fl or SMA-Cre^ERT2; R26R^DTA were administered one dose of TM for 2 consecutive days. Mice were then randomized to RT or cold exposed. Seven days later, mice were analysed for beige adipocyte formation by the following: rectal temperature (a), blood glucose levels (b), haematoxylin and eosin (H&E) staining (c), UCP1 IHC (d) and mRNA expression of beige and thermogenic markers (e). (f) SV cells were isolated from TM-induced SMA-Cre^ERT2 or SMA-Cre^ERT2; PPARγ^fl/fl mice and incubated in beige adipogenic culture conditions. Triglyceride content and mRNA expression of beige and thermogenic genes were analysed to determine adipocyte differentiation. Data are means±s.e.m. (n=4 mice per group replicated thrice). Students t-test, *P-value <0.05, SMA-Cre^ERT2; PPARγ^fl/fl or SMA-Cre^ERT2; R26R^DTA compared with control. Students t-test, #P-value <0.05, cold control compared with RT control. Students t-test, **P-value <0.01, control compared with undifferentiated SV cells. (g–k) Two-month-old Myh11-PPARγ male mice were administered one dose of TM for 2 consecutive days. Mice were then randomized to RT or cold for 1 or 2 weeks. Mice were analysed for beige adipocyte formation by the following: rectal temperature (g), blood glucose levels (h), H&E staining (i), UCP1 IHC (j) and mRNA expression of beige markers (k). Data are means±s.e.m. (n=4 mice per group replicated thrice. Scale bar, 100 μm. (l) SV cells were isolated from TM-induced Myh11Cre^ERT2; PPARγ^fl/fl mice and incubated in adipogenic conditions. Triglyceride content and mRNA expression of beige and thermogenic genes were analysed to determine adipocyte differentiation. Data are means±s.e.m. (n=4 mice per group replicated twice).