Fig. 6: DNA replication-dependent on the C-terminal non-catalytic domain of POLE1.

a–c Clone 1.6 homozygous mAID-KI cells (1.6) or clone 1.6 stably expressing myc-FLAG-Δcat - clone 12 (Δcat.12) were treated for 16 h with DMSO or dox/aux. a 5 µM ATRi was added to the indicated samples for 1 h. Western blot of the whole cell lysates is shown. 1 h aphidicolin (2 µM) treatment was used as a positive control for replication stress. b, c EdU was added for the last 30 min (b) or 2–4 h (c) of treatment, flow cytometry histograms of EdU incorporation (b) or EdU incorporation quantifications (c) are shown. Quantification is based on n = 3 independent experiments, means + SD are shown, dox-resistant population was disregarded for quantification. d–f Δcat.12 cells were treated for 16 h with DMSO or dox/aux. Ongoing replication was labeled with 10- or 40-min pulse of CldU followed by 20 min pulse of IdU and visualized using DNA fiber analysis, as described in “Methods”. Representative images (d), individual fiber lengths from a representative experiment (mean and SD) (e), and mean fiber lengths (based on n = 3 experimental repeats) and SD of the means (f), are shown. g Clone 1.6 and Δcat.12 cells were treated for 16 h with DMSO or dox/aux, followed by 10 min EdU pulse (where indicated) and iPOND isolation of proteins, associated with nascent DNA. Western blot analyses of the inputs and pulldown samples are shown. Source data are provided as a Source data file.