Lysing LRRK2 Transgenic Mouse Brains

GS LRRK2 transgenic mice (Ramonet et al. 2011 (link)) were purchased from The Jackson Laboratory (strain B6; C3-Tg [PDGFB-LRRK2*G2019S] 340D jmo/J, stock number 016575). They were housed in a specific pathogen-free facility at the Dankook University Animal Facility. The protocol was approved by the Dankook University Institutional Animal Care and Use Committee (DKU-16-035). Mice were sacrificed by the cervical dislocation. Brains were extracted and lysed in a buffer composed of ice-cold PBS with 1% Triton X-100 and 1 × protease inhibitor cocktail (Calbiochem). Each brain lysate was homogenized using a 17-gauge needle followed by incubation for 30 min at 4°C. After centrifugation at 4,000 × g for 10 min at 4°C, supernatants were transferred to a new tube and subjected to Western blot analysis.