2.7. Intracellular Rhodamine-123 Accumulation Assay

Hela cells transfected with NCL overexpression or empty vector were plated at 1 × 10⁵/well in 12-well plates and incubated for 24 h, cells were treated with or without DDP (2.5 μg/mL) for 12 h, and then, Rh123 was added at a final concentration of 10 μg/ml and incubated with cells at 37°C for 30 min. Cells were harvested and resuspended with cold PBS. The fluorescence intensity of the cells was measured at 488/575 nm using the FACS Calibur flow cytometer (Becton Dickinson, USA).