Flow cytometry analysis

A549 cells were trypsinized and aliquoted. Aliquots containing 2.5–5.0 × 10⁵ cells were stained for 30 min with the fluorescent dyes JC-1 and DCFH-DA. Thereafter, the cells were washed in PBS, centrifuged at 1200 × g, and resuspended in maintaining buffer (PBS containing 1% FBS). Cells were characterized using a FACSCalibur cell analyzer, and the data were analyzed using CellQuest software (BD Biosciences). For each run, 10⁴ events were collected. We counted at least 10,000 cells/tube for flow cytometry analysis in each experiment, respectively.