Fac scan cytometry assay kit

Cellular DNA content was determined by flow cytometry using the propidium iodide (PI)-labeling method as described previously [23 (link)]. Briefly, B16F10 cells were seeded at a density of 4 × 10⁵ cells/dish in 10 cm dishes, and the cell cycle was synchronized by the addition of double thymidine (3 mM) for 16 h. Cell cycle-synchronized cells were then washed with PBS and re-stimulated to enter the G1 phase together by the addition of fresh medium, which also contained various concentrations of CoQ₀ (0-20 μM). Cells were harvested at 24 h, and the cell cycle analysis was performed using a FAC-Scan cytometry assay kit (BD Biosciences, San Jose, CA, USA) equipped with a single argon ion laser (488 nm). The DNA content of 1×10⁴ cells/analysis was monitored using the FACScalibur system. Cell cycle profiles were analyzed with ModFit software (Verity Software House, Topsham, ME, USA).

Cellular DNA content was determined by flow cytometry using the propidium iodide (PI)-labeling method as described previously^{31 (link)}. Briefly, cells were seeded at a density of 4 × 10⁵ cells/dish in 10 cm dishes, and the cell-cycle was synchronized by the addition of double thymidine (3 mM) for 16 h. Cell-cycle-synchronized cells were then washed with PBS and re-stimulated to enter the G1 phase together by the addition of fresh medium, which also contained various concentrations of CoQ₀ (0–30 µM). Cells were harvested at 24 h, and the cell-cycle analysis was performed using a FAC-Scan cytometry assay kit (BD Biosciences, San Jose, CA, USA) equipped with a single argon ion laser (488 nm). The DNA content of 1 × 10⁴ cells/analysis was monitored using the FACS Calibur system. Cell-cycle profiles were analyzed with ModFit software (Verity Software House, Topsham, ME, USA).