Fig 2: ZFP161 Maintains Genomic Stability. a Generation and characterization of ZFP161 KO mice. A. Schematic representation of wild- type and gene-trapped alleles in the ZFP161 genomic locus. b PCR and Western blot analysis of genomic DNAs and lysates from ZFP161+/+, ZFP161+/-, and ZFP161-/- mice. F (exp) and F (obs) are expected and observed Mendelian frequencies, respectively, and n, the number of pups genotyped at three weeks after birth. Two-sided Fisher’s exact test, p = 0.02719. c Mice were treated with UV and ATR signaling was determined by Western blot of skin tissues. d–g Genomic stability was examined by measuring ?H2AX (e), metaphase spread (f), and micronucleated normochromic erythrocytes (Mn-NCE, CD71-PI + ) (g). n = 3 independent experiments. 3 mice per genotypes, 50 metaphases per mice. h–l TCGA database analysis. Tumor mRNA expression levels were obtained from TCGA for 32 paired patient samples (h). Correlation of ZFP161 expression levels with chromosomal instability levels. Number of telomeric allelic imbalances (i), frequency of LOH (j), large scale transition (k), and weighted genome integrity index (l). m Diagram showing that ZFP161 mediates the recruitment of ATRIP/ATR to repair stalled replication fork. The graphs represent mean ± S.D., two-tailed, paired t-test. Source data are provided as a Source Data file.

Fig 3: DHX9 is required for the recruitment of RAD51.a Fluorescent image (left panel) and quantification (right panel) showing the decreased formation of RAD51 foci in control U2OS cells (red) and cells knocked down for DHX9 (blue) treated with 1 µM camptothecin for 2 h then left for 2 h to recover. b DHX9 is in a common genetic pathway for the formation of RPA foci with CTIP and MRE11. RPA foci in cells knocked down using siRNA for the indicated genes are shown. Cells were treated with 1 µM camptothecin for 2 h. c Recruitment of DHX9 to foci is dependent on ATM and ATR. Wild-type cells were inhibited for ATR and ATM using VE-821 and KU55933, respectively. d, e DHX9 is in a common genetic pathway for the formation of RPA foci with ATR and ATM. RPA foci are shown for cells treated with inhibitors for ATR (VE-821) and ATM (KU55933) (d) or knocked down with siRNA against ATM and ATR (e). f Western blot showing that knockdown of DHX9 impairs Cpt induced autophosphorylation of ATR on Thr1989 and phosphorylation of Chk1 on ser 345. Quantification of n cells (as indicated) from three pooled biologically independent experiments were performed in (a–e). Means of data sets were shown to be significantly different using one-way ANOVA with Tukey’s post hoc test. (ns not significant, *p < 0.1, **p < 0.01, ***p < 0.001, and ****p < 0.0001). Error bars indicating one standard deviation are also indicated. Source data are provided as a Source Data file.

Fig 4: VMAT2 Lo mice have increased protein levels of pATR in the FC (A) at the ages of 6, 15 18, and 23 months and in the SN and (B) at the ages of 18 and 23 months (N=6/group). The upper panels in A and B show autoradiographs obtained by western blotting for pATR, ATR, and ß-actin. The lower panels in A and B show quantitative analysis of band densities.

Fig 5: VMAT2 Lo mice have increased mRNA expression levels of ATM (A) and ATR (B) in the LC and SN at the ages of 18 and 23 months, measured by qPCR (N=5/group).