Cell counting kit 8 (CCK-8) (MedChemExpress, USA) was adopted to examine the cell proliferation viability. Cells were seeded in a 96-well plate with a density of 2000 cells per well. To find out an optimal toxicity effect on the cancer cells, Anlotinib with various concentration was added 24 h after seeding in the plate of each cell lines before CCK-8 (1:10 diluted in 100 µL medium per well) was added in each plate 24, 48 and 72 h later. After incubation of the mixture for 3 hours, measurement of Formazan generated was carried out at 450 nm using a microplate reader (Molecular Devices CMax Plus, USA).
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