Xenbase Image ID: 149160

Figure 4 Replication-Dependent Repair of an ICL (A) An ICL blocks cleavage by AccI. Fifteen nanograms of pICLNm or pCtr was digested with AccI, separated on a native agarose gel, and stained with SYBR Gold. (B) ICL repair assay. At different times after addition of NPE/32P-α-dATP, pICLNm or pCtr was recovered and equal aliquots were digested with HincII (lanes 1–9) or AccI (lanes 10–18). (Note: lanes 1–9 represent a darker exposure of Figure 3B.) Twenty percent of the reaction was loaded in lanes 9 and 18. (C) At each time point shown in (B), the repair efficiency was calculated as explained in the text and graphed (red line). Extension products from the same experiment shown in Figure 4B were plotted for comparison (gray dashed line). (D) pICLNm or pCtr was replicated using NPE lacking radioactivity and optionally supplemented with p27Kip. Plasmid was recovered, digested as indicated, and examined by Southern blotting using pCtr DNA as probe. Twenty percent of the reaction was loaded in lane 13. Samples were supplemented with a 1.2 kb HindIII fragment of pCtr before extraction (loading control). Image published in: Räschle M et al. (2008) Copyright © 2008. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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