Apollo 488 in vitro kit

Before either the Cell Counting Kit-8 (CCK-8) or the EdU procedure, cells were seeded in 96-well plates at a density of 3000 cells per well for specific treatment. Cell viability was assessed using CCK-8 (Dojindo, Kumamoto, Japan). Cells were incubated with CCK-8 working solution for 3 h, after which the absorbance was measured at 450 nm using an ELx808 microplate reader (BioTeck). Relative cell viability was expressed as a proportion of specific controls.
The EdU Apollo 488 In Vitro Kit was purchased from Ribobio (Guangzhou, China) to examine cell proliferation. Medium supplemented with 20 μM EdU was added to the cells, followed by incubation for 2 h at 37 °C under 5.0% CO₂. After fixation and neutralization according to the manufacturer’s instructions, Apollo and Hoechst 33,342 staining were performed, followed by observation under an inversion fluorescence microscope (Olympus IX51 Microsystems, Japan). The percentage of EdU-positive cells was calculated from five random fields in three wells.

EdU assay were performed using Cell-Light 5-ethynyl-2’-deoxyuridine (EdU) Apollo488 in vitro Kit (Ribobio, Guangzhou, China). Briefly, GC cells were plated in 6-well plates, and incubated with 1× EdU assay buffer for 2 h. After washing, the cells were incubated with Apollo®488 staining for 30 min. The EdU(+) cells were analyzed by flow cytometry.