EdU proliferation assays (RiboBio, China) were carried out to measure cell proliferation. Cells were seeded in 96-well plates (2 × 103 cells/well) with 100 μL 10% serum-containing DMEM per well for 24 h. Cells were incubated with 50 μM EdU in serum-free DMEM for 2 h at 37°C, followed by fixing in 4% formaldehyde for 30 min on the second day. Glycine was used to neutralize formaldehyde. After permeabilizing with 0.5% TritonX-100 for 10 min at room temperature, 1×Apollo reaction cocktail (100 μl) was added to wells for 30 min. Nuclei were stained with 1×DAPI (100 μL). Cells were imaged under a fluorescence microscope (Nikon, Japan).

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