Fig. 2: Genetic screen unveils functional connections between USP37 and TRAIP.

a Biplot showing gene enrichment scores (normZ) reflecting viability in untreated conditions of WT (y-axis) and USP37 KO (x-axis) U2OS cells. b U2OS USP37 WT cells (left histogram) and USP37 KO cells (right histogram) were transfected with a CRISPR sgRNA targeting TRAIP and samples were collected 4, 7, 11, and 15 days afterwards and subjected to TRAIP sequence analysis. Percentages of unedited or edited cells at the TRAIP locus at the indicated time points are plotted. n = 3 independent experiments. Bars represent means ± SEM. c–e Clonogenic survival assays of CTRL or USP37 KO RPE-1 TP53 KO cells transduced with a LacZ control sgRNA or with a sgRNA targeting TRAIP upon treatment with camptothecin (c), ICRF-193 (d), and aphidicolin (e). n = 3 independent experiments. Bars represent means ± SEM. Half plot points indicate zero percent viability. Statistical analysis for (c–e) was performed using two-way ANOVA and Tukey test for multiple comparisons. f Representative images of RPA (green) and γH2AX (orange) staining in S-phase (magenta) positive cells. Scale bar: 20 μm. g, h Quantification of RPA (g) or γH2AX (h) foci in S-phase cells (EdU positive). Cells were treated with 20 nM camptothecin for 4 h. For (g, h), untreated: n = 1747 (CTRL, siCTRL), 1262 (CTRL, siTRAIP), 865 (USP37 KO6, siCTRL), 533 (USP37 KO6, siTRAIP), 753 (USP37 KO17, siCTRL), 458 (USP37 KO17, siTRAIP); camptothecin: n = 1885 (CTRL, siCTRL), 1315 (CTRL, siTRAIP), 838 (USP37 KO6, siCTRL), 622 (USP37 KO6, siTRAIP), 806 (USP37 KO17, siCTRL), 476 (USP37 KO17, siTRAIP). Cells were analyzed from three biological replicates. Bars represent median. Black dots indicate means in each independent experiment. Statistics indicate two-tailed Wilcoxon rank-sum tests for p values without adjustment for multiple hypothesis testing and Cohen’s d for effect sizes (γ) (see methods). Source data are provided as a Source Data file.