Sk n sh

Human neuroblastoma cells (SK-N-SH) were purchased from the Korean Cell Line Bank (Seoul, Korea). Non-tumorigenic fibroblast HS 68 (CRL-1635) cells were purchased from the American Type Culture Collection (Manassas, Virginia, USA). Cells were cultured in suitable media (SK-N-SH; RPMI-1640 media, HS 68; DMEM media) with 10% of fetal bovine serum, 100 U/mL penicillin, and 100 U/mL streptomycin at 37 °C in a humidified 5% CO₂ atmosphere. The cell passage for SK-N-SH and HS 68 subculture were within passage number 3–7.

Hela, Hek293T, SK-N-SH, SK-SH5Y, and THP-1 were obtained from the Korean Cell Line Bank (KCLB, Seoul, South Korea). Cells were grown in RPMI-1640, and supplemented with 10% fetal bovine serum (FBS) and 1% penicillin–streptomycin (HyClone, PA, USA). Rotenone was obtained from Sigma-Aldrich (MO, USA). 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) Hydrochloride was purchased from TCI chemicals (M2690, Tokyo, Japan) and dissolved in saline (0.9% NaCl, 3.75 mg/ml aliquots prepared fresh before injection). The chemicals used for neuronal cell death screening were: TNF-α (210-TA, Sigma-Aldrich, MO, USA), cycloheximide (66-81-9, MO, USA), actinomycin D (11805017, Thermo Fisher, MA, USA), cisplatin (D3371, TCI Chemical, Tokyo, Japan), paclitaxel (P1632, TCI Chemical, Tokyo, Japan), 5-fluorouracil (F0151, TCI Chemical, Tokyo, Japan), 6-hydroxydopamine hydrochloride (H4381, Sigma-Aldrich, MO, USA). The primary antibodies used were α-tubulin (sc-5286, Santa Cruz, TX, USA), Bax (2772 s, CST, MA, USA), tyrosine hydroxylase (ab6211, abcam, Cambridge, England), cleaved caspase-8 (9496 s, CST, MA, USA), cleaved caspase-9 (7237 s, CST, MA, USA), and p53 (sc-53394, Santa Cruz, TX, USA).

Human embryonic kidney (HEK293) (KCLB: 21,573), SH-SY5Y (KCLB: 22,266), and SK-N-SH (KCLB: 30,011) cells were purchased from the Korean Cell Line Bank (Seoul, South Korea). SK-N-DZ and SK-N-AS were kindly provided by Prof. Sung-Oh Huh from Hallym University, South Korea. SK-N-DZ, SK-N-AS and HEK293 cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM) (PAN^™ BIOTECH, P04-03590, Aidenbach, Germany) supplemented with 10% fetal bovine serum (FBS) (Gibco BRL, Cat. No. 10,082,147, Rockville, MD, USA) and 1% penicillin and streptomycin (Gibco BRL, Cat. No. 15,140,122, Rockville, MD, USA), while the SH-SY5Y and SK-N-SH cells were maintained in a 1:1 mixture of minimum essential medium (MEM): F12 medium supplemented with 10% FBS and 1% penicillin and streptomycin at 37 °C in a humidified atmosphere with 5% CO₂.
The transfection of plasmids and shRNA in the HEK293 cell line was performed using polyethylenimine (Polysciences, Cat. no. 25449-100, Warrington, USA). The SK-N-DZ, SK-N-AS, SH-SY5Y, and SK-N-SH cells were transfected with Lipofectamine 3000 (Thermo Fisher Scientific Cat. no. L3000001, USA) according to the manufacturer’s protocol.