Fig 1: Combination treatment does not increase SC WAT beiging more than mirabegron treatment.The protein levels of 3 markers of WAT beiging were determined in SC WAT by immunohistochemistry before and after treatment as described in Methods. The change in protein caused by mirabegron treatment (n = 13) was calculated using previously published data (15, 16); see Supplemental Figure 4. (A and B) Analysis of UCP1 expression: data from the pioglitazone (n = 12) and combination treatment groups (n = 12) were analyzed by 1-way ANOVA with data from the mirabegron treatment group as described in Methods. (C) Analysis of the change in UCP1 caused by the 3 treatments (interaction P = 0.01); the change (post-pre treatment) in protein was calculated and analyzed by 1-way ANOVA as described in Methods (**P < 0.01; *P < 0.05). (D–F) Analysis of TMEM26 expression (F: interaction P = 0.01). (G–I) Analysis of CIDEA expression (I: interaction P = 0.02). The data in (D–I) were analyzed as described for A–C. Data in C, F, and I indicate the mean ± SEM; ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05; #P < 0.1.