Western blot

For brain tissue collection, rats were decapitated and the mPFC and whole hippocampus removed. Tissues were homogenized to protein extraction of the synaptosome fraction as previously described [7, 10]. For primary cortical culture, cells were scrubbed from the wells, collected into RIPA buffer and sonicated. Homogenates were submitted to western blot analysis and the bands quantified as previously described [7, 10] (primary antibodies: rabbit anti-phospho-p70S6K, rabbit anti-p70S6K, rabbit anti-phospho-p44-42 ERK, rabbit anti-p44-42 ERK, rabbit anti-phospho-4EBP1, rabbit anti-phospho-mTOR, rabbit anti-mTOR, rabbit anti-GluA1, rabbit anti-Synapsin1, rabbit anti-PSD95, 1:1000, Cell Signaling. Secondary antibody: anti-rabbit, 1:5000, Vector Laboratories). Total levels of the respective protein or GAPDH (Cell Signaling, 1:5000) were used for loading control and normalization.