Anti th t1299

Cell lysates and SNc tissues were homogenized in RIPA lysis buffer, and protein concentration was determined by the Bradford assay (Bio-Rad, Hercules, CA, USA). A 30-μg protein aliquot of each sample was separated using standard SDS-PAGE and transferred onto a PVDF membrane (Millipore, Bedford, MA). Immunoreactive bands were detected by enhanced chemiluminescence plus detection reagent (Pierce, Rockford, IL) and analyzed using the ImageQuant™ LAS 4000 imaging system (GE Healthcare, Pittsburgh, PA, USA). The following primary antibodies were used: anti-TH (T1299, Sigma, St Louis, MO, USA), anti-p16 (sc-56330, Santa Cruz Biotechnology, USA), anti-β-actin (BM0627, Boster, Pleasanton, CA, USA), anti-LC3 (3868, Cell Signaling Technology, USA), anti-p62 (23214, Cell Signaling Technology, USA), anti-COX IV (4844, Cell Signaling Technology, USA), anti-TOM20 (42406, Cell Signaling Technology, USA), anti-Parkin (ab77924, Abcam, USA), and anti-PINK1 (ab23707, Abcam, USA).