Fig. 1: IPOND-R, a proteomics method for isolating proteins on nascent DNA at repair sites. | Nature Communications

Fig. 1: IPOND-R, a proteomics method for isolating proteins on nascent DNA at repair sites.

From: Proteomic profiling of UV damage repair patches uncovers histone chaperones with central functions in chromatin repair

Fig. 1: IPOND-R, a proteomics method for isolating proteins on nascent DNA at repair sites.

a Scheme of the IPOND-R method. b Inhibition of replication-coupled DNA synthesis measured by EdU incorporation (pulse of 10 min) in U2OS cells treated for 2 h with the replication inhibitor cocktail (Replic. inhib.). c Effect of cell pre-treatment with the replication inhibitor cocktail (Replic. inhib.) on repair-coupled DNA synthesis measured by EdU incorporation (pulse of 1 h) at sites of local UV irradiation (marked by XPA immunostaining) in U2OS cells. S phase cells present in the non-treated population were identified by pan-nuclear EdU staining and excluded from the analysis. d Western blot analysis of proteins captured by IPOND-R 1 h post UV irradiation in U2OS cells. -UV and -Biotin are used as negative controls. Experiment repeated twice with similar results. Dot plots: mean ± SD from n cells scored in 3 independent experiments (the mean of each experiment is shown as a gray triangle). Statistical significance is calculated with a two-sided Student’s t test with Welch’s correction. Scale bars, 10 μm.

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