(A and B) mRNA expression of the indicated genes in BAT, iWAT, and eWAT (A) and Western blot of UCP1 and mitochondrial proteins (B) from male C57BL/6 mice at thermoneutrality after tail vein injections of AAV-GFP or AAV-PM20D1. Mice were placed into thermoneutrality (30°C) at 6 weeks old, injected with virus at 7 weeks old, and high fat diet (HFD) was started 7 days post injection. Mice were maintained at 30°C for the duration of the experiment. For (A) and (B), data are from 47 days post injection. For (A), n=8/group, mean ± SEM, * p<0.05. For (B), n=4–5/group.
(C) Chromatogram at m/z = 428 from plasma of male C57BL/6 mice after tail vein injection of AAV-GFP or AAV-PM20D1. For (C), mice were 7 weeks old at the time of injection, high fat diet (HFD) was started 7 days post injection, and mice were maintained at room temperature for the duration of the experiment. The comparative metabolomics was performed on plasma harvested 54 days post injection. n=4/group, * p<0.05.
(D) Chemical structure of N-oleoyl phenylalanine (C18:1-Phe).
(E and F) MS/MS spectra (E) and retention time (F) of endogenous (top) or synthetic (bottom) C18:1-Phe.
See also Table S3.