Fig. 2: RPA-marked inherited genomic lesions occur at telomeres.
From: RPA shields inherited DNA lesions for post-mitotic DNA synthesis
a Asynchronously growing U-2 OS cells were treated for 24h as indicated. Cells were stained for RPA and TRF2, and cell cycle staging to identify G1 cells was performed by QIBC. Representative images of individual G1 cells are shown. b Quantification of the average number of RPA foci co-localizing with TRF2 in G1 in different treatment conditions from n = 3 independent samples with n1 = 499, n2 = 495, n3 = 476 (Control), n1 = 367, n2 = 345, n3 = 278 (APH), n1 = 902, n2 = 1046, n3 = 968 (ATRi), n1 = 959, n2 = 1042, n3 = 1044 (APH + ATRi) cells in G1 per sample. Individual average values and means ± SD are show. P-values were determined by two-tailed unpaired t-test; **p < 0.01 (exact p-values are p = 0.0094, p = 0.005, and p = 0.0066, respectively). c Quantification of the percentage of RPA foci co-localizing with TRF2 in G1 corresponding to b. Individual average values and means ± SD are shown. d Quantification of the percentage of TRF2 foci co-localizing with RPA in G1 corresponding to b. Individual average values and means ± SD are shown. P-values were determined by two-tailed unpaired t-test; **p < 0.01 (exact p-values are p = 0.0066 and p = 0.0044, respectively). e Native (non-denaturing) FISH-IF to detect ssDNA at telomeres in U-2 OS cells. A telomeric FISH probe (Tel C) was used to detect single-stranded G-rich telomeric sequences. Tel C foci counts were quantified by QIBC and are plotted in a cell cycle-resolved manner. At least 1000 cells per condition were analyzed. f Representative images of G1 cells from native FISH-IF staining using the telomeric Tel C probe and RPA. Scale bars: 10 μm. A. U., arbitrary units. Source data are provided as a Source Data file.
