Doxorubicin Cytotoxicity Assay

Cells in the logarithmic growth period were cultured in a 96‐well plate at a density of 2 × 10⁵ cell/mL in a 0.1 mL volume for 6 h. Doxorubicin of variable concentrations (0.1, 0.2, 0.3, 0.4 and 0.5 μg/mL) was added to the experiment group, while the control group was cultured with the same volume of culture medium in a 5% CO₂ incubator at 37℃ for 24 h. A minimum of 3 parallel wells were set in each group. Cell viability was detected using a WST‐1 kit (Roche Applied Science, Upper Bavaria, Germany). One hour before each measurement, 100 µL Dulbecco’s modified Eagle’s medium and 10 µL WST‐1 solution were added successively, the absorbance (A) value at the wavelength of 450 nm was documented using a microplate reader. The inhibitory concentration IC₅₀ was calculated to compare the resistance.¹⁷ (link)