Mouse Brain Homogenization and Protein Extraction

Mouse brain homogenates were prepared as described in our earlier study56 (link). Briefly, mice brain tissues were snap frozen in liquid nitrogen when harvested and the wet weight of the tissues (in mg) was determined using an electronic balance. The entire left hemisphere of the brain was digested with ice-cold 1x RIPA lysis buffer (Cell Signaling Technology) containing detergents (1% Nonidet P40 and 1% sodium deoxycholate) together with the protease inhibitor cocktail tablet (Roche). Lysates were homogenized using a hand held motorized pestle (Sigma-Aldrich) on ice. Tissue lysates were collected for protein quantification using BCA analysis (ThermoFischer Scientific)