Cell counting kit cck8

FGSCs (2000 cells per well) were plated in 96-well plates, treated for 48 h with GANT61 (HY-13901, MedChemExpress, USA), and then measured using a Cell Counting Kit (CCK-8, Transgen BioTECH, China). In addition, 2000 FGSCs were seeded in 6 cm plates and cultured for 7 days under GANT61 treatment. Crystal violet was used to stain and count the number of cellular colonies.

The experiment was conducted as described elsewhere with minor modifications [28 (link)]. In brief, cell Counting Kit (CCK)-8 (TransGen Biotech, Beijing, China) was used to determine cell proliferation according to the manufacturer's instructions. HepG2 cells were transfected with either mutant or wild-type S-gene recombinant pcDNA3.1(-)/myc-His A plasmids. The cells were seeded onto 96-well plates at 2000 cells/well after 12 h of the transfection,. At 12, 24, 36, 48 and 60 h, the culture medium in each well was replaced with 200 ml fresh medium mixed with 10 ml CCK-8 solution. The absorbance of each well was measured using a microplate reader (BioTek, USA) at 450 nm. The experiment was independently performed for three times.

GCs were spread on a 96-well plate at 2000 cells per well and treated with cisplatin at different concentrations (0, 5, 10, 15 and 20 μM) for 24 h or with 20 μM cisplatin for different durations (0 h, 3 h, 6 h, 12 h, to 24 h). A Cell Counting Kit (CCK8, Transgen BioTECH, Beijing, China) was used to detect cell proliferation activity. Primary GC were isolated from ovarian follicles as described previously, and cell density reached 70%-80% after 48 h of culture. After treatment with lentiviral suspension for 48 h, cells were seeded into 6-well plates (1000 cells per well) for 5 days for clone formation experiments and into 96-well plates (4000 cells per well) for 48 h for EdU staining (KGA331, KeyGEN BioTECH, China). Crystal violet (G1062, Solarbio, China) was used to stain and count the number of colonies.