Trypan blue staining assay kit

Cell viability was evaluated by utilizing trypan blue staining assay kit (Beyotime, China). After transfection, primary cardiomyocytes and H9C2 cells were cultivated in a 6-well plate (1 × 10⁵ cells per well) (Thermo Fisher Scientific, USA) at 37 °C for 24 h. Subsequently, PBS and the kit solution were separately used to wash and fix the collected cells, which were counted with a microscope (Nikon, Japan). Finally, cell viability was calculated based on the formula: (the number of viable cells/the number of total cells) × 100%.

Cell viability was assessed using trypan blue staining assay kit (Beyotime Biotechnology, China) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltertrazolium bromide tetrazolium (MTT) assay (Sigma-Aldrich). For trypan blue staining, after relevant transfection, GH3 cells were seeded into a 6-well plate (Thermo Fisher Scientific, USA) with 1 × 10⁵ cells per well and cultured at 37°C for 24 h. Then, cells were collected, washed with phosphate-buffered saline (PBS), stained using the kit solution, and counted under a microscope (Nikon, Japan). Cell viability (%) was calculated by number of viable cells / number of total cells × 100%.
For the MTT assay, after relevant transfection, GH3 cells were seeded into a 96-well plate (Thermo Fisher Scientific) with 1 × 10⁴ cells per well and cultured at 37°C for 24 h. Then, 20 μL MTT solution (2.5 mg/mL in PBS) was added into the medium of each well and the plate was incubated at 37°C for 4 h. Subsequently, the MTT mixture was removed and 150 μL dimethyl sulfoxide (DMSO) was added to dissolve formazan. After that, the plate was agitated on a shaker for 15 min. The absorbance of each well at 570 nm was recorded using a microplate reader (Bio-Tek Instrument, USA).