Clone formation assay
The
clone formation experiment was used to evaluate the effect of NCTD on
the cloning ability of A549 cells. A549 cells were cultured in a 6-well
microplate (1000 cells / well) in 2 ml of 10% fetal calf serum and 5%
CO2 humidified atmosphere at 37 ° C overnight. The cells were then
treated with the prepared drug working solution NCTD on a 6-well plate
for 7 days, and then stained with 0.1% crystal violet (Sigma, St.
Louis, Mo, USA) for 15 minutes. A digital camera (Canon, EOS350D, Tokyo,
Japan) was used to obtain images of the number of colonies.