Fig. 3. Repair of HBV lesions in human nuclear extracts.

a A simplified schematic depicting the four fragments in unrepaired RrcDNA digested by AatII/PsiI. b NA-RrcDNA (lanes 1–5) or RrcDNA (lanes 6–11) was incubated with human nuclear extracts in a time course assay. Repair products at various time points were digested with AatII/PsiI and subsequently resolved by UREA-PAGE gel. Repair of the plus-strand Pa fragment was monitored by Southern blot as in Fig. 2d. “*” indicates extended Pa fragments that reach the 5′ terminus of Pa. c Repair of the plus-strand RNA primer-containing Pb fragment was monitored by Southern blot as described in Fig. 2e. Traces with dashed blue and orange lines denote unprocessed (blue arrow head) and processed product (orange arrow head) of Pb fragments. d–e Repair of the minus-strand Ma and Mb fragments were monitored by Southern blot as in Fig. 2f, g. f Removal of the biotin-containing flap of Ma was detected by streptavidin blot as in Fig. 2h. g The repair efficiency of plus- and minus-strands is calculated from c and d and plotted as in Fig. 2i. (−) and (+) repair denote repair of minus and plus-strands, respectively. All experiments were repeated three times, and each individual measurement is plotted. The lines connect the average value of three measurements at each time point. Statistical analyses between the repair efficiencies of the minus strand at each time point are performed by two-stage step-up t test method from Graphpad Prism. P values are 0.00001, 0.0003, 0.0003, and 0.000005 at indicated time points (*). Source data are provided as a Source Data file.