Xenbase Image ID: 156415

Figure 3. XPF‐ERCC1 mutant complexes are active in nucleotide excision repair (NER) Schematic representation of unscheduled DNA synthesis (UDS) during NER on a UV‐treated template in high‐speed supernatant (HSS) egg extract. Mock‐depleted, XPF‐ERCC1‐depleted (ΔXE), and XPF‐ERCC1‐depleted extracts complemented with wild‐type (XEWT) or mutant XPF‐ERCC1 (XEMUT) were incubated with untreated or UV‐treated plasmids for 2 h at room temperature in the presence of 32P‐α‐dCTP. Reaction products were isolated, linearized with HincII, and separated on a 0.8% agarose gel. The DNA was visualized by autoradiography to show incorporation of 32P‐α‐dCTP during UDS (upper panel) and stained with SYBR gold for total DNA (lower panel). The incorporation of 32P‐α‐dCTP was quantified, the background signal from non‐damaged plasmid was subtracted, and the signal for the mock depletion condition was set to 100% to normalize the data. Error bars represent s.e.m. of three independent experiments. **P = 0.003, ***P = 0.0004, paired t‐test comparing all conditions to the mock. All non‐marked conditions did not show a statistical difference from the mock condition. Source data are available online for this figure. Image published in: Klein Douwel D et al. (2017) © 2017 The Authors. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

Return to previous page