Sk ov 3 human ovarian adenocarcinoma cell line

4T1 murine mammary carcinoma cell line, MCF-7 human breast adenocarcinoma cell line, HepG2 human hepatocellular carcinoma cell line, NIH/3T3 murine fibroblast cell line, NDF normal human dermal fibroblast cell line, MDA-MB-231 human breast adenocarcinoma cell line, PC12 rat pheochromocytoma cell line, HeLa human cervical adenocarcinoma cell line, and SKOV3 human ovarian adenocarcinoma cell line (purchased from American Type Culture Collection, ATCC) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% antibiotics (penicillin and streptomycin) and placed in a humid CO₂ incubator providing an atmosphere containing 5% CO₂ at 37 °C.
To detect the intracellular ROS aroused by HSN, cells were seeded in confocal cell culture dishes at a density of 1 × 10⁴ cells per dish. After culture in incubator for 12 h, HSN (final concentration: [pTBCB] = 50 µg mL⁻¹) or PBS were added to the cells and incubated with cells for 24 h. Thereafter, cells were gently washed three times with fresh PBS to remove excess nanoparticles, and then fixed with 4% paraformaldehyde. After fixation, cells were successively stained with DAPI and DCF-DA. Then cells were imaged under a LSM800 confocal microscope.