Dna cell proliferation kit

Cell proliferation was tested using EdU (5-ethynyl-2’-deoxyuridine) DNA Cell Proliferation Kit (RiboBio, Guangzhou, China) and CCK8 kit (Doindo, Japan). After required treatment, WI-38 cells were incubated with 50 nM EdU for another 2 h. The proliferating cells were fixed with 4% paraformaldehyde and incubated with Apollo Dye Solution. Cell nuclei were stained with Hoechst 33342. For CCK8 assay, transfected cells were plated in 96-well plates at a density of 5 × 10³ cells per well. After required treatment, CCK8 reagent was added to each well to incubate for another 2 h. The absorbance was measured at 450 nm wavelength using a microplate reader (Thermo Fisher, NewYork, USA).

3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium-bromide assay (MTT) was used to determined cell viability. The cells were cultured in 96-well plates at a density of 1 × 103 cells per well for 5 days and counted daily. The absorbance value was determined at 490 nm, and the cell proliferation rates were measured by cell growth curve.
EdU (5-ethynyl-2′-deoxyuridine) DNA Cell Proliferation Kit (RiboBio, Guangzhou, China) was used for cell proliferation analysis. Cells were grown on coverslips and incubated with 50 mM EdU for 2 h. After washing and fixing, the cells were incubated with Apollo Dye Solution to stain the proliferating cells. Counterstain was performed using Hoechst (1:1000). Images were randomly captured from six fields (Nikon 80i, Nikon Corporation, Tokyo, Japan).
Cells were plated in 6-well plates and scratched with sterile 10 μl pipette tip to create a wound. After washing, medium without serum was added to the well. Images were taken at 72 h from at least six fields.
The cell migration ability was determined by Matrigel-coated transwell inserts. Cells were plated to Matrigel-coated inserts and allowed to invade through the Transwell plates. The cells attached to the other side were fixed, stained and counted. The number of invading cells was averaged from at least six randomly chosen fields.