Figure 4.
DPY30 Deficiency Results in Increase in DNA Damage and Impairs DNA Repair
(A) Immunoblotting of γ-H2AX and H3 in Vav-Cre; Dpy30F/+ and Vav-Cre; Dpy30F/− FL and Lin− BM cells from pIpC-injected Mx1-Cre; Dpy30F/+ and Mx1-Cre; Dpy30F/− mice. Bottom, quantification from five animals each.
(B) Scheme for DNA damage assays and the mouse genotype legend for (C–E). LSK cells sorted from tamoxifen-treated Lin− BM were irradiated for assays at different time points. n = 3 mice each.
(C) Relative Dpy30 mRNA levels were determined by qRT-PCR and normalized to Actb.
(D) γ-H2AX foci numbers per cell from 20 to 30 LSK cells each. Representative images are in Figure S5F.
(E) Comet assay on LSK cells. We used four classes of cell morphology to show increasing DNA damage severity, and quantified (bottom) the percentages of each class before and after irradiation.
(F) Assay scheme and the mouse genotype legend for (G). Tamoxifen-treated Lin− BM were irradiated, followed by FACS assays gated on LSK cells at different time points. n = 3 mice each.
(G) FACS analyses for phosphorylated Kap-1 in LSK cells. Left, representative plots. Right, quantification based on the demarcation of the vertical line shown in the left plots.
Data are shown as mean ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, by two-tailed Student's t test. Scale bars, 10 μm. See also Figure S5.