Sk n mc

The human neuroblastoma cell line SK-N-MC was obtained from Korean Cell Line Bank (Seoul, Korea). The antibodies of p-Akt (Thr308), p-Akt (Ser473), Akt, mTOR, Caveolin-1, Flotillin-2, p-Tau (Ser396), Tau, p-NF-κB p65 (Ser536), NF-κB p65, Lamin A/C, CBP and β-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The antibodies of p-mTOR (Ser2448), p-p70S6K1 (Thr389) and p70S6K1, were acquired from Cell Signaling Technology (Beverly, MA, USA). Aβ, BACE1, HIF-1α and GPR40 antibodies were obtained from Abcam (Cambridge, MA, USA). The C99 antibody was purchased from EMD Millipore (Darmstadt, Germany). Horse radish peroxidase (HRP)-conjugated IgG was obtained from Jackson Immunoresearch (West Groove, PA, USA). PA, BSA, GW9508, ionomycin, PF4708671, LY294002 and rapamycin were purchased from Sigma Chemical Company (St. Louis, MO, USA). The Akt inhibitor, GW1100 and SN50 used here were purchased from Calbiochem (La Jolla, CA, USA). siRNAs for GPR40, GPR120, APP, BACE1, HIF-1α and non-targeting were obtained from Dharmacon (Lafayette, CO, USA).

SK-N-BE(2)-M17 (male), SH-SY5Y (female), and Neuro-2A mouse neuroblastoma cells were purchased from American Type Culture Collection (Manassas, VA, USA), and IMR-32 (male), SK-N-SH (female) and SK-N-MC (female) human neuroblastoma cells were obtained from Korean Cell Line Bank (Seoul, Korea). All cells except IMR-32 were cultured in DMEM supplemented with L-glutamine (300 mg/ml), 25 mM HEPES, 25 mM NaHCO₃, 10% heat-inactivated FBS and antibiotic-antimycotic agents at 37 °C in a 5% CO₂ incubator. IMR-32 cells were grown in RPMI media supplemented with L-glutamine (300 mg/ml), 25 mM HEPES, 25 mM NaHCO₃, 10% heat-inactivated FBS and antibiotic-antimycotic agents. Before treatment with CFZ, Cis, or other drugs, cells were refreshed with the culture media and stabilized for 3 h.