SOD1 Is Essential for the Viability of DT40 Cells and Nuclear SOD1 Functions as a Guardian of Genomic DNA
Figure 1
Generation of cells expressing human SOD1 from the tet-off promoter. (a) Schematic representation of the generation of DT40 hSOD1 cells. (b) Schematic representation of the SOD1 genomic locus and configuration of the targeted locus. (c) Southern blot analysis for the confirmation of the status of the SOD1 wild-type (WT), WT + human SOD1 (+/+), heterozygous (+/−), and homozygous (−/−) cells, using NdeI digested genomic DNA and the probe indicated in (b). (d) Dox-dependent disappearance of human SOD1, as confirmed by western blot analysis, with α-tubulin used as the loading control. (e) Growth curves of indicated genotype cells. Indicated mutant cells were cultured in the presence or absence of 1 g/mL Dox. Dox had no effect on the growth or viability of wild-type cells.
