Apollo dye solution

The proliferative capacity of lentivirus-infected LUAD cells transfected with or without SP3 expression plasmid or vector control for 48 h was analyzed according to the manufacturer’s instructions of the Cell-lightTM EdU DNA cell proliferation kit (Cat#C10310-3, Ribobio). Briefly, LUAD cells (1×10⁴) were seeded into 96-well plates. On the second day, the cells were incubated with 10 μM EdU for 2 h. After fixation with 4% paraformaldehyde (Cat#P1110, Solarbio), the cells were stained with Apollo Dye Solution. The nuclei of the cells were stained with Hoechst 33342. Image acquisition was performed using an Olympus SZH10 Stereo microscope (Olympus, Tokyo, Japan).

EdU experiment was conducted with Cell‐Light EdU DNA Cell Proliferation Kit (RiboBio, Guangzhou, China). 3 × 10⁴ cells were plated into 24‐well plates and incubated for 48 hours. After incubation in 300‐µL EdU for 2 hoours, HCT116 and SW620 cells were fixed by 4% paraformaldehyde (Solarbio) and stained by Apollo Dye Solution. DAPI was applied to dye nucleic acid. Images were collected via an inverted fluorescence microscope (Olympus) and proportion of EdU‐positive cells was examined.