Separate brains from 8 HAP (4 male, 4 female) and 8 LAP (4 male, 4 female) mice were rapidly harvested by a blinded researcher and the DS, as a whole, was dissected bilaterally and homogenized. Tissue collection was completed in the morning to midday and samples were deidentified until tissue was fully processed and data was prepared for analysis. Flash frozen brain lysates were prepared in 1 mL of 9 M urea (CHEBI: 16199) in 20 mM HEPES, pH 8.0 (CHEBI: 46756), 1x cOmpleteTM protease inhibitor cocktail (Roche Diagnostics cat. 11836153001). Tissues were homogenized using a BeadBugTM 6 (Benchmark scientific Cat No: D1036, 3mm zirconium beads Cat No: D1032–30, 10 rounds of 30 × 30 sec,4 °C). Samples were next sonicated in 1.5 mL Micro Tubes (TPX Plastic for Sonication from Diagende Inc.) using a Bioruptor® sonication system (Diagenode Inc. USA, North America cat number B01020001) with 30 sec/30 sec on/off cycles for 15 minutes in a water bath at 4 °C. After subsequent centrifugation at 12,000 rpm for 20 min, protein concentrations were determined by Bradford protein assay (BioRad
