Updated FTP location. A 2-mg frozen section of hypothalamus from a single mouse was homogenized in 200 µL of denaturing solution(7 M urea, 2 M thiourea, 20 mM dithiothreitol) containing cOmplete Mini Protease Inhibitor Cocktail (Roche). Peptide extracts equivalent to 135 µg of tissue were analyzed by LC-MS/MS.