Figure 4. Effect of WWL113 on UCP1 expression and cellular respiration.
A: UCP1 protein expression in differentiated Ucp1-luciferase brown adipocytes treated with WWL113 for 5 days. Rosiglitazone (0.5 μM) served as positive control.
B: Expression of thermogenic genes in differentiated Ucp1-luciferase brown adipocytes treated with WWL113 (10 μM) for 5 days. Rosiglitazone (0.5 μM) was served as positive control (n=3). * P <0.05, *** P < 0.001 vs. control.
C: Ucp1 mRNA expression in primary differentiated brown adipocytes treated with WWL113 (10 μM) or rosiglitazone (5 μM) for 24 or 48 hours (n=3). *** P < 0.001 vs. control.
D: Ucp1 mRNA expression in primary differentiated brown adipocytes treated with WWL113 (10 μM) or rosiglitazone (5 μM) for 48 hr. Norepinephrine (0.1 μM) was added 8 hr prior to harvest (n=3). ** P <0.01, *** P <0.001 vs. control.
E: Total and uncoupled (oligomycin-insensitive) respiration of differentiated brown adipocytes (5 × 105 cells/sample) treated with WWL113 (10 μM) in the presence or absence of norepinephrine (0.1 μM) (n=3–4). * P <0.05 vs. control.
F: Ucp1 mRNA expression in differentiated brown adipocytes treated with vehicle or WWL113 (10 μM) for 24 hr with or without 30 min pre-treatment with a PPARα-selective antagonist (GW6471; 3 μM) and/or a PPARγ-selective antagonist (GW9662; 10 μM) (n=3). *** P < 0.001 vs. control. ### P< 0.001 vs. WWL113-treated cells.
G. Ucp1 mRNA expression in differentiated brown adipocytes treated with vehicle, WWL113 (10 μM), a PPARα-selective agonist (GW9578), or the combination for 24 hr (n=4). * P < 0.05, *** P < 0.001.