Coronal brain sections (16 μm) were probed as published (8,18,26). We used specific oligonucleotides for AgRP, CART, fatty acid synthase (FAS), NPY, and POMC detection (Supplementary Table 1). Anatomical regions were analyzed using the rat brain atlas (27). Sections were scanned, and the hybridization signal was quantified by densitometry using ImageJ-1.33 (National Institutes of Health, Bethesda, MD) (8,18,26). We used between 16 and 20 sections for each animal (4 to 5 slides with four sections per slide). The mean of these 16–20 values was used as the densitometry value for each animal.
