For cell proliferation assay, the transfected cells were seeded into 96-well plates at a density of 2000 cells per well. At 0, 24, 48, 72 and 96 h after seeding, cell viability was measured by the cell counting kit-8 (CCK-8) system (Beyotime, China) according to the manufacturer’s instructions. Briefly, each well was added with 10 μl CCK- 8 solution, and the plate was then incubated at 37 °C for 1 h in dark. Absorbance at 460 nm of each well was measured using a microplate reader (Tecan, Switzerland).
Cck 8 system
Manufactured by Beyotime
Sourced in China
The CCK-8 system is a colorimetric assay kit used to measure cell viability and cytotoxicity. It utilizes the water-soluble tetrazolium salt WST-8 to produce a water-soluble formazan dye upon reduction by dehydrogenases in viable cells. The amount of formazan dye generated is directly proportional to the number of living cells, allowing for quantitative analysis of cell proliferation and cytotoxicity.
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3 protocols using cck 8 system
1
Cell Proliferation Assay Using CCK-8
2
Apigenin Inhibits ACC-2 Cell Proliferation
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Cultured ACC-2 cells were trypsinised with 0.25% trypsin (100biotech Co., Ltd). Cell proliferation was measured using the CCK-8 system (Beyotime Institute of Biotechnology, Nanjing, China), according to the manufacturer's instructions. Briefly, 3×103 ACC-2 cells were seeded into 96-well culture plates and treated with either DMEM, or 10, 40 or 160 µM apigenin (Selleckchem, Houston, TX, USA) The cells were cultured in serum-free medium (100biotech Co., Ltd.) at 37°C for 1, 2, 3, 4 or 5 days. A total of 10 µl CCK-8 reagent was added to each well, and incubated at 37°C for 3 h, and the absorbance was measured at 450 nm. Optical density (OD) was calculated as follows: OD = ODcell − ODblank.
3
Cell Proliferation Assays for Cancer
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We performed the Cell Counting Kit-8 (CCK-8) assay to examine cell proliferation. The transfected cells were seeded into a 96-well plate at a density of 1000 cells/well. We measured cell viability through the CCK-8 system (Beyotime, China). For EdU assay, 97H and G2 cells were stained using a EdU assay kit (RiboBio, China) following to its instructions. After that, a fluorescence microscope was used to take pictures, with 3 fields randomly selected for each slide. Lastly, the number of EdU-positive cells was counted and quantified. Regarding the colony formation experiment, about 1000-2000 cells were seeded in each well of the 6-well plate, allowing cells to grow until visible colonies were formed.
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