Cell viability assay kit

The HCC cells were plated at 5000/well in 96-well plates and treated with vehicle or THZ1 following a 10-fold serial dilution. Five days later, cell viability was measured using a cell viability assay kit (#ab232855, Abcam, Cambridge, MA, USA). Briefly, a crystal violet staining assay was performed to determine cell viability according to the manufacturer’s instruction. After the removal of the culture medium and gentle washing with a washing solution, a crystal violet staining solution (50 μL) was added to each well and incubated for 20 min at room temperature. After thorough washing, a solubilization solution was added and incubated again for 20 min at room temperature on a benchtop rocker. The optical density (O.D.) of each well was measured at 595 nm. Data were represented as the percentage of viable (attached) cells calculated against the values of the cells treated with vehicle.

The cell viability was measured following the procedure of a previous study [19 (link)]. A cell viability assay kit (Abcam, Cambridge, UK) was utilized according to the instructions of manufacturer.

The MCF-7/Dox (Doxorubicin resistant) cells were seeded in a 96-well plate at a density of 2 × 10⁴ cells/100 µL per well. The cells were allowed to attach overnight and then treated with different concentrations of DHA, doxorubicin, alone, or in combination in fresh media. After 48 h of incubation, the culture medium was removed, and the cells were washed twice with PBS. The adherent cells were then stained with crystal violet staining solution (Cell Viability Assay Kit (ab232855), abcam, Boston, MA, USA) for 20 min, followed by washing and air-drying. The solubilization solution was added, and the absorbance was measured at 570 nm using a microplate reader as per the manufacturer’s instructions.