Mtt cell cytotoxicity assay kit

Neonatal myocytes were isolated from 1-day-old SD rats and cultured as described previously [25 ]. Briefly, cells were maintained in DMEM supplemented with 10 % FBS at 37 °C in a humidified incubator with 95 % air and 5 % CO₂. Cells viability was measured with an MTT cell Cytotoxicity Assay kit (Beyotime, Shanghai, China). The absorbance was read at 570 nm. Six independent experiments were performed.

To determine the effects of Hsp70 inhibitor VER155008 on Vero E6 cell viability, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was applied. Briefly, Vero E6 cells were grown on 96-well culture plates at a density of 5 × 10³/well and were incubated for 24 h. After initial period of incubation, cells were treated with various concentrations of VER155008 for another 24 h. Cell viability was assessed using the MTT Cell Cytotoxicity Assay Kit (Beyotime, Beijing, China) as the manufacturer’s recommendations.

Cisplatin cytotoxicity was investigated by a MTT Cell Cytotoxicity Assay Kit (Beyotime, Shanghai, China). MKN-7/DDP and HGC-27/DDP cells (10,000 cells per well) were placed into 96-well plates overnight and then incubated with a series of concentrations (0–20 μg/mL) of cisplatin for 48 h. Then, the cells were incubated with MTT reagent for 4 h and then incubated with formazan dissolving solution. Subsequently, the output of the absorbance at OD570 nm was determined with a microplate reader (Bio-Rad, Hercules, CA, USA). The half maximal inhibitory concentration (IC₅₀) of cisplatin was estimated according to the survival curve.