Fig. 5: Disruption of the WRN-RPA complex leads to persistence of G4s in the cell.

a Analysis of G4 structures by immunofluorescence and an anti-DNA G-quadruplex antibody (clone BG4) in WS cells or WS cells expressing Flag-WRN^wt or Flag-WRN^6A plasmid. Cells were treated as indicated. The graph quantifies total BG4 nuclear staining per nucleus from two independent experiments (at least 80 nuclei/repeat). Data points from inhibited cells have a black border. Bars represent mean ± S.E. (ns = not significant; *P < 0.05; **P < 0.01: Two-tailed Mann-Whitney test for paired samples). Representative images are provided. Scale bar = 20 µm. b SIRF analysis of the localisation of BG4 at EdU-labelled nascent DNA by in situ PLA in WS cells expressing Flag-WRN^wt or Flag-WRN^6A. To mark nascent DNA at stalled forks, cells were treated with EdU 8 min before being treated with HU. The graph quantifies total BG4 SIRF spots per nucleus from two independent experiments (at least 80 nuclei/repeat). Bars represent mean ± S.E. (ns = not significant; ****P < 0.0001; Kruskal-Wallis with Dunn’s test). Representative images are provided. Scale bar = 20 µm. c Analysis of G4 structures detection as in a) in WS cells expressing Flag-WRN^wt or Flag-WRN^6A. Cells were treated as indicated and recovered with or without WRN inhibitor (WRNi). The graph shows quantification of total BG4 nuclear staining per nucleus from two independent experiments (at least 80 nuclei/repeat). Data points from inhibited cells have a black border. Bars represent mean ± S.E. (ns = not significant; **P < 0.01; ****P < 0.0001; Two-tailed Mann-Whitney test for paired samples). Representative images are provided. Scale bar = 20 µm. Source data are provided as a Source Data file.