M7449

SH-SY5Y, a human neuroblastoma cell line, and HT22, an immortalized mouse hippocampal neuronal cell line, were cultured as previously described.^{55 (link)} Cells were transfected with cDNA for Crif1 (gifted from Dr. Shong (Chungnam University, Korea)), Sp1 (hMU003598; provided from Korea Human Gene Bank, Medical Genomics Research Center, KRIBB, Korea), Sp1 K16A or Mito-DsRed using Lipofectamine and Plus reagents (Invitrogen, Carlsbad, CA, USA) and Crif1 siRNA (Santa Cruz Biotechnology, Santa Cruz, CA, USA) using RNAiMax reagents (Invitrogen) for 48 h according to the manufacturer's instructions. Cells were grown to 60% confluency and treated with vehicle (dimethyl sulfoxide) or Aβ_1-42 (American Peptide, Sunnyvale, CA, USA), Aβ_42–1 (Bachem, Bubendorf, Switzerland) and various reagents; MG132 (10 μM; M7449, Sigma-Aldrich, St. Louis, MO, USA), 3-MA (2 mM; M9281, Sigma-Aldrich), bafilomycin (5 nM; B1793, Sigma-Aldrich), NAC (1 mM; A7250, Sigma-Aldrich), apocynin (10 μM; A10809, Sigma-Aldrich), DPI (10 μM; D2926, Sigma-Aldrich), and H₂O₂ (216763, Sigma-Aldrich).

NPC cells (SUNE1, CNE-2, 5–8 F and CNE-1), human normal nasopharyngeal epithelial cells NP69 and human embryonic kidney (HEK) 293 T cells were purchased from Shanghai Huzhen Biotechnology Co., Ltd (Shanghai, China). SUNE1 and HEK293T cells were cultured in DMEM (A4192101, Gibco, USA), while other cells were cultured in RPMI-1640 medium (A4192301, Gibco), added with 10% foetal bovine serum (FBS; 16140071, Thermo Fisher Scientific, USA). All cells were kept in a humid incubator with 5% CO₂ at 37°C. For cell treatment, cycloheximide (CHX; 5 µM; 5087390001, Sigma-Aldrich, St. Louis, MO, USA), MG132 (10 Μm; M7449, Sigma-Aldrich), actinomycin D (4 μg/Ml; SBR00013, Sigma-Aldrich) and p65 inhibitor (parthenolide; PT; 5 μmol/L; HY-N0141, MedChemExpress, New Jersey, USA), were purchased.