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. Author manuscript; available in PMC: 2017 Oct 1.
Published in final edited form as: Nat Genet. 2017 Feb 13;49(4):537–549. doi: 10.1038/ng.3790

Figure 7. DONSON patient cells have spontaneous defects in replication fork progression that result in DNA damage.

Figure 7

(a) Complementation of patient-derived fibroblasts with WT DONSON. Fibroblasts derived from DONSON patients P2, P6 and P9 were infected with retroviruses encoding either WT DONSON or an empty vector. DONSON expression was determined by immunoblotting. A non-specific cross-reactive protein represents a loading control. (b) Expression of WT DONSON in patient fibroblasts rescues elevated levels of spontaneous DNA damage. The percentage of cells from (a) with 53BP1/γH2AX foci was quantified by immunostaining (n=3). (c) DNA fibre analysis of complemented DONSON patient fibroblasts pulsed with CldU and IdU. Fork asymmetry was quantified. Plot indicates ratios of left/right fork track lengths of bidirectional replication forks. The red lines denote median ratios. (n=3). (d) The percentage of stalled forks and new origins from cells in (c) was quantified (n=3). Ongoing forks are shown in (Supplementary Fig. 19).