Quick cell proliferation colorimetric assay kit

The effects of Newcastle disease virus (NDV) AF2240 on the proliferation of MDA-MB231 cell line were assessed using Quick Cell Proliferation Colorimetric Assay Kit (BioVision, USA). Briefly, the cells were grown in a 25 cm² until their confluent. Trypsinized cells were counted as previously described [26 (link)]. Cells were plated in 96-multiwell microtiter flat bottom plate (Nunc, Denmark) at a density of 1 × 10⁴ cells/mL in a final volume of 100 μL/well in culture medium and then incubated for 24 hours to allow confluent. Old media were removed and replaced with a new media containing the graded concentration of the virus (twofold dilution). Some wells were left untreated but treated with PBS to serve as control. Triplicate experiments were run before the cells were incubated for 24 hours in a humidified 5% CO₂ incubator at 37°C. After the end of this treatment period in incubation, 10 μL of the lyophilized WST reagent was added to each well and further incubated for 3 hours in a standard culture conditions. The reaction mixture was stopped by adding 10 μL of the stop solution into each well. The cultured cells were then shaked thoroughly for 1 minute. Finally, microtiter reader at 420–480 nm wavelengths measured the absorbance of the treated and untreated samples.

MDM cells were seeded in a 96 well plate and transfected with IL-4pDNA/GFP-NPs for 24 hours. Non-treated cultures served as the control. After 24 hours, cell viability was assayed using the Quick Cell Proliferation Colorimetric Assay Kit (BioVision, Milpitas, CA) according to the manufacturer’s protocol. Samples were analyzed with the FlexStation 3 Multi-Mode Microplate Reader (Molecular Devices, San Jose, CA) at 450 nm, and the data collected was normalized to the controls. The experiment was repeated in triplicate.