Fig. 6: ATAD5 depletion increases cell death and genomic instability in cells and mice.

a, b U2OS cells transfected with ATAD5 siRNA under the Noco-APH condition were treated with 2 mM HU for 6 h, plated in the absence of drug treatments, incubated for 2 weeks, and examined to detect surviving colonies by methylene blue staining. a Representative images of colony staining. b The viability of the surviving colonies was quantified. Error bars represent standard deviation of the mean (n = 3). Statistical analysis: two-tailed Student’s t test; *p < 0.05. c, d U2OS cells transfected with ATAD5 siRNA under the Noco-APH condition were treated with HU for 6 h and subjected to metaphase spreading. c Representative images of DAPI stained chromosomes are shown. Arrows point to chromosomal breaks. d The number of chromosomal breaks per cell was plotted. Statistical analysis: two-tailed Student’s t test; *p < 0.05. e U2OS-TetOn-ATAD5 cell lines expressing wild-type or two ATAD5 mutants were treated with doxycycline and transfected with ATAD5 siRNA under the Noco-APH condition. Cells were then treated with HU for 6 h and subjected to metaphase spreading. Statistical analysis: two-tailed Student’s t test; *p < 0.05. n.s., not significant. f Atad5 wild-type or mutant mice were analyzed by micronucleus analysis. Error bars represent standard deviation of the mean (n = 8). Statistical analysis: two-way analysis of variance, ***p < 0.001. g The model described in the text.