G2019s tg mice

The cell lysates were centrifuged at 16,000 × g for 10 min at 4 °C to remove insoluble material. The supernatants were precleared, then immunoprecipitated with anti-p53 or anti-LRRK2 (MJFF2, Abcam) at 4 °C for 8 h, and further incubated with protein-A agarose (Pierce, Rockford, IL,USA) for 24 h. The antibody-protein complexes were subjected to Western blot analysis using the indicated antibodies.
Cell nuclei were fractionated using a Nuclear/Cytosol Fractionation kit (K266-100, Bioscience, Milpitas, CA, USA) as suggested by the manufacturer, after the indicated treatment.
G2019S TG mice (#009609, Jackson Laboratory, Bar Harbor, ME, USA) and normal control littermates were sacrificed by cervical dislocation. Brain tissues were disrupted using a Dounce Homogenizer (10 strokes) and lysed by passing the extracts through a 22-gauge needle five times. The brain lysates were centrifuged at 16,000 × g for 30 min at 4 °C to remove insoluble material, and supernatants were used for further Western blot analysis.
The immunoprecipitated antibody-protein complexes, cell or brain lysates, or fractionated samples were subjected to Western blot analysis using the indicated antibodies.

G2019S TG mice were purchased from The Jackson Laboratory (strain B6; C3-Tg [PDGFB-LRRK2*G2019S] 340D jmo/J, stock number 016575) (Ramonet et al. 2011 (link)) and were housed in a specific pathogen-free facility at the Dankook University Animal Facility with the approval of the Dankook University Institutional Animal Care and Use Committee (DKU-16-035). G2019S mice and littermates were sacrificed by cervical dislocation. Brains were removed from the skull and lysed by ice-cold PBS with 1% Triton X-100 and 1Xprotease inhibitor cocktail (Calbiochem). Brain lysates were homogenized using a 17-gauge needle, incubated for 30 min at 4°C, and centrifuged at 4,000×g for 10 min at 4°C. Each supernatant was transferred to new tube and analyzed by western blotting.