Countess automated cell counter tc10 automated cell counter

To measure cell proliferation, the cells were seeded in 24-well plates at a density of 2.0 × 104 cells per well. The cells were cultured for 2 days and cell medium was renewed daily. Where indicated, the cells were treated with Rotenone (24 h at the indicated concentrations), then trypsinized every 24 h and quantified using the Countess automated cell counter TC10 automated cell counter (Bio-Rad). The time-course experiments were repeated three times.

To measure cell proliferation, the cells were seeded in 24-well plates at a density of 2.0 × 10⁴ cells per well. The cells were cultured for 3 days and cell medium was renewed daily. Afterwards, the cells were trypsinized every 24 h and quantified using the Countess automated cell counter TC10 automated cell counter (BIO-RAD). The time-course experiments were repeated three times using cells derived from three individual mice per each group. Where indicated, cells were incubated with BrdU (1.5 μg ml⁻¹; Sigma-Aldrich) in accordance with the manufacturer’s protocol. BrdU-labeled cells were detected by immunostaining using rat anti-BrdU antibody (Oxford Biotechnology; 1:500), followed by the specific secondary biotinylated goat anti-rat antibody (1:300; Jackson Immunoresearch) and mounted with the Prolong Gold Anti-fade reagent. The slides were analyzed by using a Leica SP8 confocal laser scanning microscope and quantified by using CellProfiler.