- Cells are seeded and cultured in a medium containing 10% fetal bovine serum .
- Cells were seeded and incubated for 72 h prior to addition of the BrdU solution.
Cells were seeded at a density of 50 000 cells per well in a six-well plate and maintained in media containing 10% fetal bovine serum. At the end of the experiment, cell number was determined using trypan blue exclusion. Cell proliferation assays were carried out using the bromodeoxyuridine (BrdU) cell proliferation assay kit (BioVision, Milpitas, CA, USA) according to the manufacturer's protocol. Briefly, 10 000 cells/well were seeded into a 96-well plate and incubated for 72 h prior to addition of the BrdU solution. BrdU incorporation was measured by absorbance at 450 nm. For clonogenic assay, cells were seeded onto 60 mm culture dishes at the density of 1000 cells/well. Cultures were maintained for 9–12 days in complete culture media until colonies appeared. The colonies were fixed with 4% paraformaldehyde and stained with 0.5% crystal violet. The plates were photographed and the numbers of visible colonies were counted. Full paper
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