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. 2017 May 4;2(9):e91981. doi: 10.1172/jci.insight.91981

Figure 9. BH4 directly activates brown adipocytes NO-dependently.

Figure 9

(A and B) mRNA levels of brown adipose tissue–related (BAT-related) genes by quantitative RT-PCR (A; n = 4–5) and protein expression levels of UCP1 (B; n = 3) in differentiated brown adipocytes on day 7 after incubated with or without 10 or 100 μM tetrahydrobiopterin (BH4) for 24 hours. (C–E) Changes of oxygen consumption rate (OCR) (C); values of OCR related to basal respiration, ATP production, proton leak, and maximum respiration (D); and coupling and uncoupling rate (E) in the differentiated brown adipocytes on day 7 incubated with or without 100 μM BH4 for 24 hours (n = 7–8). (F) NO production in the differentiated brown adipocytes on day 7 incubated with or without 10 or 100 μM BH4 for 24 hours (n = 5). (G) mRNA level of Ucp1 in the presence of the 100 μM nitric oxide synthase (NOS) inhibitor (L-NAME) in the differentiated brown adipocytes on day 7 incubated with or without 100 μM BH4 for 24 hours (n = 5–6). Values are mean ± SEM. Statistical analysis was performed by Student’s t test (C–E and G). *P < 0.05, **P < 0.01, NS vs. control group (C–E) or vs. values without BH4 (G). In addition, statistical analysis was performed by 2-way ANOVA (C) or 1-way ANOVA with Tukey post-hoc test (A, B, and F). *P < 0.05, **P < 0.01 vs. control group.