Measuring Cell Proliferation Dynamics

Cell proliferation was measured by CCK-8 assay and EdU incorporation assay. For CCK-8 assay, A172 and U251 cells were seeded in 96-well plates, and cell viability was eveluated using the Cell Counting Kit-8 kit (C0038, Beyotime) according to the manufacturer’s instructions. Cell viability was expressed as the percentage of control group. The EdU incorporation assay was conducted as previously reported [18 (link)]. In brief, proliferating A172 and U251 cells cultured in 24-well plates were examined using the Click-iT Edu Proliferation Assay Kit (C10337, ThermoFisher Scientific) according to the manufacturer’s instructions. The Edu incorporation was analyzed by the flow cytometry (Accuri C6, BD). Each group included five replicates and three independent experiments were performed.