96 well plate reader

Primary chondrocytes were plated in triplicate in 96-well plates (1000 cells/well) in appropriate growth medium. The cell proliferation/viability assay was performed by following the WST-1 kit instructions (Sigma_Aldrich, USA). Briefly, at the indicated time points, cell medium was replaced with 90 µL fresh growth medium supplemented with 10 µL WST-1 reagents, followed by incubation at 37 °C for 1 h. OD absorbance values were measured at 490 nm using a 96-well plate reader (Bio-Rad, USA).

The CellTiter 96 Aqueous One Solution Cell Proliferation assay system (Promega, Madison, WI, USA), which is a colorimetric approach for the detection of the cytotoxic effect of a compound, was used to measure cell viability according to the manufacturer’s protocol. Ava5 cells were seeded in 96-well plates at a density of 5 × 10³ cells per well and treated with SFN at indicated concentrations for 3 days. The absorbance values were detected at 490 nm using a 96-well plate reader (Bio-Rad, Hertfordshire, UK).

The complex 1 (STR2) was dissolved in minimum quantity of DMSO, diluted in culture medium and used to treat HepG2 hepatocellular carcinoma cells, in 96 well culture plates, at a concentration of the complex ranging from 1 to 10 μM for a period of 24 h. Cisplatin was used as the reference drug, and DMSO was used as the solvent control. A miniaturized viability assay using 3-(4,5-di-methyithiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) was carried out according to the method described by Mosman (1983). The cells were assayed by the addition of 20 μL of the MTT solution (5 βmg mL-1) prepared in phosphate-buffered saline (PBS). The plates were wrapped in aluminum foil and incubated for 4 h at 37 °C. The purple formazan product was dissolved by the addition of 100 μL of 100% DMSO to each well. The absorbance was monitored at 570 nm (measurement) and 630 nm (reference) using a 96 well plate reader (Bio-Rad, Hercules, CA, USA). Data were collected for triplicates and used to calculate the respective means. The percentage inhibition was calculated from this data using the formula:

The IC₅₀ value was determined as the concentration of the complex/cisplatin that is required to decrease the absorbance to half that of the control.

The viability of PoAgNPs-treated MDA-MB-231 cells was measured using the MTT assay [25 (link)]. For this, the human adenocarcinoma breast cancer cells (MDA-MB-231) at 1.5 × 10⁶ cells/well were seeded in 96-well plates to treat with different concentrations of PoAgNPs (10–100 μg/mL) and incubated in a CO₂ incubator for 24 h. After incubation, 20 μL of MTT solution (5 mg/mL in phosphate-buffered saline) was added. After 4 h, the purple formazan product was dissolved by adding 100 μL of 100% DMSO to each well. The absorbance was monitored at 570 nm (measurement) and 630 nm using a 96-well plate reader (Bio-Rad, Hercules, CA, USA). All the assays were done in triplicate, and the results were given in mean ± SD
$Percentage of inhibition=\frac{(OD value of control-OD value of Sample}{OD value of control}\times 100$

Cells were seeded in 6- or 12-well plates. The next day, cells were treated with L-ASP, enzalutamide, or both for four hours and subjected to 4 Gy of irradiation. Cells were grown for 10–14 days to allow for colony formation. Cells were then fixed in 100% methanol for 20 min and stained with 5% crystal violet in methanol for 5 min. Plates were washed extensively in water before solubilizing using 30% acetic acid solution in water. Solubilized samples were analyzed by Bio-Rad 96-well plate reader using the 595 nm filter. Absorbencies were normalized to the control per experiment.