Figure 7
mtDNA replication is required for increase in the number of mt-nucleoids. (a) Effect of ddC treatment on mtDNA and nuclear DNA replication. Cells were treated with or without 100 μM ddC for 3 h, and subsequently incubated with 20 μM EdU for 1 h. Position of the nucleus (white dotted line) is shown in each image. Scale bar, 10 μm. (b) Effect of ddC treatment on cell cycle progression. Cells were treated with 100 μM ddC and time-lapse imaged at 1-h intervals. Error bars indicate standard deviation (ncells = 20 for each phase). Statistical significance was examined by Student’s t-test. N.S., not significant (p = 0.4422). Scale bar, 10 μm. (c) Effect of ddC treatment on regulation of mt-nucleoid number. Fucci2 cells treated with ddC were time-lapse imaged at 4-h intervals. Error bars indicate standard deviation (ncells = 10 for each bar). (d) Effect of aphidicolin treatment on mtDNA and nuclear DNA replication. Cells were treated with 15 μM aphidicolin (Aphidicolin) or 1:4000 DMSO (Control) for 10 h, and subsequently incubated with 20 μM EdU for 1 h. Position of the nucleus (white dotted line) is shown in each image. Scale bar, 10 μm. (e) Effect of aphidicolin treatment on regulation of mt-nucleoid number. The increase in the number of mt-nucleoids within 4 h was investigated during the phase in which the color of the nucleus changed from orange to green, in Fucci2 cells treated with 15 μM aphidicolin or 1:4000 DMSO for 10 h. Statistical significance was examined by Student’s t-test. N.S., not significant (p = 0.4325). Error bars indicate standard deviation (ncells = 10 for each condition).
