P nf κb p65 ser536

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).

Febuxostat (Febutaz) was purchased from Sun Pharma, Sikkim, India, and allopurinol (Zyloric) was purchased from GlaxoSmithKline, Mumbai, India. The drugs were suspended in carboxymethyl cellulose (0.5% CMC). The kits for TNF-α and IL-6 were purchased from RayBiotech Inc., Norcross, GA. The CK-MB kit was purchased from Logotech Private Limited, India.
The primary antibodies for caspase-3 (#9662S), Bcl-2 (#2876S), and β-actin (#4967S) were procured from Cell Signaling Technology, MA, USA, and those for Bax (#7480), NF-κBp65 (#109), p-NF-κBp65 (Ser536) (#101752), JNK (#572), p-JNK (#6254) (Thr 183/Tyr 185), ERK1/2 (#135900), p-ERK1/2 (#16982) (Thr 202/Tyr 204), and p-p38 (#17852) (Thr 180/Tyr 182) as well as anti-rabbit (#2004) and anti-goat (#2020) secondary antibodies were procured from Santa Cruz, California, USA. Antibody for p38 (#197348) was purchased from Abcam Technologies, USA.