Sk n sh human neuroblastoma cells

SK-N-SH human neuroblastoma cells obtained from American Type Culture Collection (HTB-11, Manassas, VA) were grown in complete F12-Dulbecco’s Modified Eagle Medium (ThermoFisher Scientific) as previously described^{15 (link)}. Complete media contained 10% fetal bovine serum (ThermoFisher Scientific) and 1% penicillin–streptomycin antibiotic (ThermoFisher Scientific). Cellular extracts used in the CaMKII assay were centrifuged at 15,000 rpm for 10 min at 4 °C. Protein concentrations of the extracts were determined by Bradford assay using the Protein Assay Kit II (Bio-Rad, Hercules, CA).

All chemicals, solvents, and reagents used for extraction, fractionation, in vitro biological, and in vivo toxicity evaluations were purchased from Sigma Aldrich (St. Louis, MO, USA), Merck (Branchburg, NJ, USA), Systerm (Shah Alam, Malaysia), Nacalai Tesque (Kyoto, Japan), Molecular Probes (Eugene, OR. USA), Invitrogen (Waltham, MA, USA), Fisher Scientific (Waltham, MA, USA), San Francisco Bay Brand (Newark, NJ, USA) and Sera (Grapevine, TX, USA) and used without further purification. The BV-2 mouse-based microglial and SK-N-SH human neuroblastoma cells of American Type Culture Collection (ATCC, Rockville, MD, USA) were provided by Prof. Dr. Johnson Stanslas of the Pharmacotherapeutics Lab, Faculty of Health and Sciences, Universiti Putra Malaysia.