N-labeled mice were dissected and flash frozen in liquid nitrogen. We next prepared P2 membrane fractions from the frozen cortical tissue. All cortices were homogenized in cold sucrose buffer supplemented with protease inhibitor cocktail and a BCA was perform to assess the protein concentration of each sample. Following quantification,+/+
mouse homogenate. Homogenates were then centrifuged at low speed to pellet nuclei and cell debris and the supernatant was then centrifuged at high speed to obtain the membrane fraction . Membrane pellets were solubilized in cold RIPA buffer for 1 h at 4C and clarified by centrifugation . Proteins were then extracted using methanol/chloroform precipitation and stored at −80Membrane fraction were resuspended with 100 μl of 8 M urea for 30 min. Then 100 μl of 0.