Wst 1 reagent

NPCs were transduced with SOCS3-expressing or control lentivirus and cultured for 48 h. Then NPCs were reseeded at 5×10⁴ cells/well in poly-D-lysine-coated 96-well plates, and cell viability was analyzed by the WST-1 assay on days 1, 3, and 5. The WST-1 reagent (10 μl, Beyotime) was added to each well (100 μl) and incubated at 37°C for 2 h. The absorbance at 450 nm was determined with 690 nm as the reference using an Epoch microplate spectrophotometer (BioTek Instruments, USA).

Differentiated THP-1 cells were treated with histamine, 2-PE, or cetirizine at appropriate concentrations for 24 h in 96-well plates (5 × 10³ cells/well); cells treated with DMSO were included as a negative control. After treatment, WST-1 reagent (Beyotime, Shanghai, China) was added (10 μL/well) and the cells were incubated for 2 h at 37°C. The formazan dye produced from WST-1 by viable cells was quantified by measuring the absorbance at 450 nm using the Epoch^TM 2 microplate spectrophotometer. The absorbance at 650 nm was also measured as the reference wavelength. Cell viability was calculated according to the manufacturer’s instructions.

Cells were seeded in 96-well plates at a density of 1 × 10⁴ cells/well, and were treated with various chemotherapeutic agents for either 48 or 72 h. Cell viability was measured with either AlamarBlue^® solution (Invitrogen) or WST-1 reagent (Beyotime, Shanghai, China) following the manufacturer’s instructions. Chemotherapeutic agents and chemicals dimethylsulfoxide (DMSO), bicalutamide, abiraterone, docetaxel, etoposide and hydrogen peroxide (H₂O₂) were purchased from Sigma. Charcoal dextran-stripped serum (CDSS) was obtained from Gemini (West Sacramento, CA, USA) and Enzalutamide was obtained from Selleck Chemicals (Houston, TX, USA).