Figure 6
Abnormalities of ROS production by and reduced size of Drp1 cells. (a) Measurement of total cellular ROS. Vector and Drp1 cells were exposed to CM-H2DCFDA as previously described4, 7 and analyzed by flow cytometry. Nearly identical results were observed on three separate occasions (not shown). The ratios of the mean fluorescence are indicated above the panels. (b) Measurement of mitochondrial superoxide production using MitoSOX staining in Vector and Drp1 cells. (c) ROS production by myc+/+ (TGR1) fibroblasts and myc−/− rat fibroblasts. Cells were plated and stained with CM-H2DCFDA and MitoSOX as described in panels (a) and (b). (d) Cell size determinations. Vector and Drp1 cells were plated onto 100-mm tissue culture dishes, and grown for two days to allow the maximal proliferative rates to be achieved. The cells from subconfluent confluent plates were then harvested by trypsinization, stained with trypan blue and the diameters of all viable cells were determined on a Beckman-Coulter Vi Cell analyzer. At least five independent cell size determinations were performed at different times with a minimal of 3000 cells per determination. The mean cell diameter±1 S.E. is shown
