EdU-Based Proliferation Assay

The EdU Apollo567 In Vitro kit (Ribobio, China) was used to assess cell proliferation. Cells transfected with miRNA mimics for 24 h were seeded into 6-well plates (2 × 10⁵ cells/well). After culture for 12 h cultivated, the cells were transfected for 24 h and were cultivated for 40 h. The cells were then incubated in EdU working solution for 2 h, fixed with 4% paraformaldehyde, permeabilized, washed, and stained with 1× Apollo solution and 1× Hoechst33342 solution, according to the manufacturer’s instructions. Results were analyzed from microphotographs taken using a fluorescence microscope.