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. Author manuscript; available in PMC: 2024 Jan 22.
Published in final edited form as: Science. 2023 Sep 22;381(6664):eadi3448. doi: 10.1126/science.adi3448

Fig. 5. DONSON is required for DNA replication and CMG assembly in mammalian cells.

Fig. 5.

(A) Immunoblot of synchronized DONSON-AID2 HCT116 cells. TCE, total cell extract; Asy, asynchronous; G1, G1-arrest; R, release, hours. (B) G1-synchronized cells fail to progress into S phase after 5-Ph-IAA depletion of DONSON. (C) DONSON depletion prevents DNA synthesis. EdU pulse-labeling at 17 hours post-release. Flow cytometry plots in panels (B) and (C), representative of n = 4 and n = 3 experiments, respectively, are quantified in figs. S16D and E, respectively. (D, E). DONSON is required for CDC45 and GINS recruitment to chromatin. (D) Immunoblots, soluble extract (Sol.) and chromatin-bound proteins (Chrom.) 17 hours post release. (E) Quantification, normalized to loading control (soluble, α-Tubulin; chromatin, Histone H2B) and wild-type protein levels. Datapoints, n = 3 experiments. Mean ± SEM.