Edu experiment kit

EdU assay was performed according to the manufacturer's instructions of the EdU experiment kit (RiboBio). Transfected cells were inoculated into a 96‐well plate at a density of 10,000 cells per well. The proportion of EdU‐positive cells was the proportion of proliferating cells. Images were acquired by fluorescence microscopy (Olympus). The results were obtained from three biological replicates. Each experiment was repeated in triplicate.

The assay was performed according to the manufacturer’s instructions of the EdU experiment kit (RiboBio, Guangzhou, China). EdU (5-ethynyl-20-deoxyuridine) is a nucleoside analog of thymine that can be integrated into DNA during DNA synthesis. The cells were inoculated into a 96-well plate at a density of 6,000 cells per well. After 24 h, 100 μl of 0.1% EdU reagent was added to each well for 2 h, followed by fixation with 4% formaldehyde at room temperature for 30 min. After treatment with 0.5% Triton for 10 min, the cells were incubated with 1×Apollo^® reagent for 30 min. Finally, Hoechst 33342 was added to counterstain cells at room temperature for 30 min. The proportion of EdU-bound cells was the proportion of proliferating cells. The results were obtained from three independent repeated experiments.