Beyoclicktmedu 488 cell proliferation kit

Infected COLO205 and SW480 cells were seeded in 24-well plates at a density of 1 × 10⁵ cells/well for 24 h. After treatment with NP (10^–6 M) or DMSO (n = 3 per group) for 24 h, the cells were treated with 5-ethynyl-2′-deoxyuridine (EdU) and counted using the BeyoClickTMEdU-488 cell proliferation kit (Beyotime, Hangzhou, China) according to the manufacturer’s instructions. The cell nuclei were stained with 4′,6-diamidino-2-phenylindole. They were then visualized under a fluorescence microscope, photographed, and the number of positive cells (green fluorescence) were analyzed using the ImageJ software.

CCK-8 assay was performed to determine the number of living cells during cell proliferation. The HepG2 cells were inoculated in 96-well plates at a density of 5 × 10³/well. After transfection for 24, 48, 72 or 96 h, 10 μL CCK-8 reagent (Transgen Biotech Co., LTD) was added to each well and incubated at 37 ℃ for 1 h. Measure the absorbance at 450 nm according to the instructions.
EdU (5-Ethynyl-2′-deoxyuridine) assay was performed to detect HepG2 cells in the dividing phase (S phase) and analyze HepG2 cell proliferation using BeyoClick^TMEdU-488 Cell Proliferation Kit (Beyotime). The density of HepG2 cells was adjusted, and 5 × 10³ per well were spread in a 24-well plate. After transfection for 24 h, EDU reagent was mixed into the culture medium for 12 h. After fluorescence staining, the cell density was observed under a fluorescence microscope.

EdU assay was conducted using BeyoClick^TM EdU-488 Cell Proliferation Kit (Beyotime, China). Transfected cells were cultured in medium containing 3μg/ml cisplatin (Sigma-Aldrich, USA). EdU labeling was conducted at the appropriate treated time. Briefly, cells were exposed to 10μM EdU for 2 hours at 37°C. Afterwards, cells were fixed with 4% paraformaldehyde followed by permeabilized with 0.3% Triton X-100 (Solarbio, China, diluted by 1×PBS). Click reaction solution was added to wells for 30 minutes' incubation at room temperature in the dark. Subsequently, cells were stained with Hoechst 33342 to label nucleus. Images were captured using inverted fluorescence microscope (Olympus, Japan). EdU-positive cells' counting and merged photographs' generation was completed using Photoshop CC 2019 software.