Cck 8

Cell clone formation assay: The cells were transfected in 6-well plates (500 cells per well) and fixed with 4% paraformaldehyde. The cells were stained with crystal violet, and the number of clones was counted as >50 cells/group.
CCK-8 assay: After transfection and other treatments in a 96-well plate (about 1,500 cells per well, 3 parallel wells), a volume of 10 μl of CCK-8 (BBI Life Sciences, China) was mixed with 90 μl of culture medium at 0, 24, 48, 72, and 96 h after transfection, and incubated for an additional 2 h at 37°C. A microplate spectrophotometer (Biotek, USA) was utilized, and the absorbance was measured at 450 nm.

Neuronal viability was assessed with a cell viability counting kit CCK-8 purchased from BBI Life Sciences (Shanghai, China). Briefly, cells after treatment were collected following the manufacturer’s protocol. Cell suspension at 100 μl was transferred to a 96-well plate at a final concentration of 500,000 cells/ml. Cells were incubated with 10 μl CCK-8 solution from the kit per well for 4 h and assessed with a PerkinElmer spectrometer (PerkinElmer, USA) at the absorbance of 450 nm.