Assays

Proteomics: brain tissues coming from both human PFC and the 4 different mouse models were prepared as follows. Tissues were grinded with a biomasher using 350 µL of MeOH:H2O (4:1). Protein pellets were resuspended in 200 µL Laemmli buffer (10% SDS, Tris 1M pH 6.8, glycerol) then centrifuged at 11.135 rpm at 4°C for 5 minutes. Protein concentration was determined using DC assay (BioRad, Hercules, CA, USA) according to the manufacturer’s instructions. 100 µg of protein lysate for each sample were ...

Phosphoproteomics: starting from the protein extracts from the global proteomics experiments, proteases inhibitors (Sigma, P8340) and phosphatases inhibitors (final concentration in Na3VO4 = 1 mM) were added to all samples. Protein concentration was determined using RC-DC assay (BioRad, Hercules, CA, USA) according to the manufacturer’s instructions. 250 µg of proteins for each sample were reduced and alkylated prior to an in-house optimized single-pot, solid-phase-enhanced sample preparation ...

Library-Matched Single Shot (LMSS) method Mice were sacrificed by cervical dislocation and brains were subsequently extracted and put into cold phosphate buffered saline (PBS). The ventricular walls were laid bare by removing the dorsal ventricular wall and all tissue above it, as well as the choroid plexus. Brains were then snap-frozen on dry ice and cut into 100 μm sections on a cryostat (Leica CM1000S). The medial (MEZ) and lateral ventricular (SEZ) walls were then manually dissected under a ...