Dimethyl sulfoxide (dmso)

The MTT assay was used to detect cell proliferation. SH-SY5Y cells were seeded in 96-well plates. Three days after transfection with IL-3 siRNA, the cells were subjected to OGD for 24 hours and then incubated in the serum-free medium containing 0.5 mg/mL MTT (Beyotime) for 4 hours at 37°C. Subsequently, 100 μL dimethyl sulfoxide (Biosharp) was added to each well. The optical density at 570 nm was measured using a Thermo Scientific Microplate Reader.

Hieff Trans Liposomal Transfection Reagent and PAGE Gel Quick Preparation Kit (12.5%) were purchased from Yeasen (Shanghai, China). Penicillin-streptomycin solution (100 ×), RIPA lysis buffer, and crystal violet were sourced from Beyotime (Shanghai, China). Fetal bovine serum (FBS) and RPMI-1640 medium were obtained from Bio-Channel (Nanjing, China). TRIzol reagent and dimethyl sulfoxide were purchased from Biosharp (Hefei, China). 5-Azacytidine was acquired from Selleck (Houston, United States of America). Paraformaldehyde was obtained from Servicebio (Wuhan, China). Cell counting kit-8 (CCK-8) was sourced from topscience (Shanghai, China). Nitrocellulose filter (NC) membranes were purchased from PALL (New York, United States of America). TMEM100 and β-actin primers were procured from Tsingke (Beijing, China). TMEM100 monoclonal antibodies were purchased from Proteintech (Wuhan, China). Human monoclonal antibodies against extracellular regulated kinase 1/2 (ERK1/2), phosphorylated (p-) ERK1/2, the c-Jun N-terminal kinase (JNK), phosphorylated (p-)JNK, p38, phosphorylated (p-) p38, goat anti-rabbit horse radish peroxidase (HRP) IgG, goat anti-mouse HRP IgG, and GAPDH were purchased from Zen Bioscience (Chengdu, China).

Roswell Park Memorial Institute (RPMI)-1640 medium and fetal bovine serum (FBS) were obtained from Gibco (Grand Island, NY, USA). Shikonin, N-acetyl-L-cysteine (NAC), and necrostatin-1 (Nec-1) were procured from Sigma (St. Louis, MO, USA). Pan-caspase inhibitor z-VAD-fmk was obtained from Tocris (Bristol, UK). The Caspase-8 Fluorescence Metric Assay and Caspase-3 Fluorescence Metric Assay Kits were bought from KeyGEN BioTECH (Nanjing, China). The Reactive Oxygen Species Detection Kit and Protein Concentration Assay Kit were obtained from Beyotime Institute of Biotechnology (Wuhan, China). Annexin V Apoptosis Detection Kit I was purchased from BD Pharmingen (San Jose, CA, USA). Dimethyl sulfoxide (DMSO) was procured from Biosharp (Hefei, China). Radioimmunoprecipitation assay (RIPA) buffer, containing protease and phosphatase inhibitors, was obtained from Sigma (St. Louis, MO, USA). Anti-RIPK1, anti-RIPK3, anti-MLKL, anti-β-actin, and anti-GAPDH antibodies were obtained from Cell Signaling (Danvers, MA, USA) and diluted 1:1,000 in TBST. Anti-rabbit IgG HRP-linked antibody and anti-mouse IgG HRP-linked antibody were also obtained from Cell Signaling (Danvers, MA, USA) and diluted 1:5,000 in TBST. The enhanced chemiluminescence (ECL) western blotting detection reagent was purchased from Wanleibio (Shanghai, China). Cell Counting Kit-8 was purchased from Dojindo (Kumamoto, Japan)

Niclosamide was obtained from Sigma-Aldrich (St. Louis, MO, USA). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT; also known as thiazolyl blue and methylthiazolyldiphenyl tetrazolium bromide) was purchased from Sigma-Aldrich. The Annexin V-FITC Apoptosis Detection kit was purchased from Vazyme Biotech (Nanjing, China). Dimethyl sulfoxide (DMSO) was obtained from BioSharp (Hefei, China). The primary antibodies, rabbit polyclonal anti-Notch1 (ab27526), rabbit polyclonal anti-Notch2 (ab8926), rabbit polyclonal to anti-Notch3 (ab23426) and rabbit polyclonal anti-Hey1 (ab22614) were purchased from Abcam (Cambridge, UK). Mouse anti-β-actin monoclonal antibody (TA-09) was purchased from ZSGB-BIO (Beijing, China). The secondary antibodies, peroxidase-conjugated AffiniPure goat anti-rabbit IgG (ZB-2301) and peroxidase-conjugated AffiniPure goat anti-mouse IgG (ZB-2305), were purchased from ZSGB-BIO.