Figure 3

Depletion of WRNIP1 increases the frequency of chromosome breaks upon PhenDC3 treatment. (a) Quantification of γH2AX intensity in shCtrl and shWRNIP1 carrying HeLa cells. Where indicated, cells were treated with G4 ligand PhenDC3 (10 µM). Cells were immunostained at different time points after PhDC3-treatment. γH2AX intensity was measured in 100 nuclei per sample. (b) Representative images of untreated and PhenDC3-treated cells from (a). Scale bars, 50 μm. (c) Chromosome aberration assay. Quantification of chromosome breaks per chromosome spread in shCtrl and shWRNIP1HeLa cells. Where indicated, cells were treated with G4 ligand PhenDC3 (10 µM). After 12 h, samples were treated with colchicine for 6 h to arrest the cells in M phase. Chromosome aberration was counted in 40 chromosome spreads per sample. (d) Representative images from (c). Scale bars, 10 μm. Error bars represent SEM of 3 biological replicates on graphs (B) and (D). p-values were obtained by ANOVA (Origin Pro) (*P < 0.05, **P < 0.01, and ***P < 0.001). Statistical data is provided in Table S4.