Fig 1: NEAT1 promoted NEDD4L mediated PINK1 degradation and impaired PINK1-dependent autophagy. (A) NEAT1 promoted interaction of NEDD4L and PINK1 in SH-SY5Y cells. SH-SY5Y cells were transfected with NEAT1 and cell lysates were immunoprecipitated by anti-NEDD4L antibody and subjected to western blotting using anti-PINK1 antibody. (B) NEAT1 regulated ubiquitination of PINK1 in SH-SY5Y cells. NEAT1 or knockdown plasmids accompanied by HA-ubiquitin and PINK1-Flag were delivered into SH-SY5Y cells. The cell lysates were subjected to immunoprecipitation assay followed by western blotting with an anti-Ubiquitin antibody. (C) NEAT1 regulated PINK1-dependent autophagy. SH-SY5Y cells transfected with NEAT1 and shNEAT1 were subjected to western blot using PINK1, P62, OPTN, and LC3I/II. (D) NEAT1 regulated amyloid-β accumulation via autophagy signaling in N2A-APPsw cells. The levels of amyloid-β42 (Aβ42) in N2A-APPsw cells transduced with lentivirus encoding shNEAT1 and co-transfected with siRNA against PINK1, OPTN, LC3, or control siRNA were measured by amyloid-β ELISA (n = 4). **p < 0.01 (E) Representative immunoblot bands for amyloid-β proteins in N2A-APPsw cell lysates with above treatment, and β-actin served as a loading control. The bar graph presents the quantification of bands for amyloid-β relative to β-actin. n = 4 independent experiments of each group. **p < 0.01.