Cell Counting Kit-8

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	- Cells were incubated for 3 h at 37 °C after addition of CCK-8 reagent

Protocol tips
- Cells were incubated for 3 h at 37 °C after addition of CCK-8 reagent

Publication protocol

Cells were seeded into a 96-well plate with 5000 cells in each well. Cell proliferative ability was detected by using the CCK-8 kit according to the kit instructions (Beyotime Institute of Biotechnology, Shanghai, China) 24, 48 and 72 h after culture and transfection. Ten microliters of CCK-8 reagent were added to each well, and plates were stored at 37°C in the incubator for 3 h. After that, the optical density (OD) at 450 nm was detected with a multifunctional microplate reader (Thermo Fisher Scientific Inc., Waltham, MA). The concentration of dimethyl sulfoxide (DMSO) in each well was no more than 0.1%, and four replicate wells were set up. The above processes were repeated 3 times, and the average value was collected. The influence of HAS-2 silencing on cell proliferation was calculated on the basis of the OD value of each well. With the cell viability (%) of each group at different time points as the detection index, cell viability (%) = (experimental group OD - blank control group OD)/(control group OD - blank control group OD) × 100%.

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