Fig. 7: ssDNA digestion by dna2-AC, is less processive and sensitive to DNA structures.

a Impact of titrated RPA on the digestion of internally labeled 562-nt ssDNA (5 nM) by Dna2 or dna2-AC respectively. For quantification, mean values ± s.d. from three independent experiments were plotted. b Digestion by titrated Dna2 and dna2-AC, on 3′-labeled 140-nt 5′-overhanging ssDNA (5 nM) (hairpin-containing) without or with RPA. The experiments were repeated three times. c Wide-field image in DNA Curtains experiments of dna2-AC (magenta puncta) digesting the RPA–ssDNA complexes (green). d Dna2 recruitment (%) for different experimental conditions (Supplementary Table 1). Independent DNA Curtains experiments were repeated: n = 3 for Exp. 1, 2, 4, 8, 9, 10. Error bars, mean ± s.d. e Boxplot of τ1 for different experimental conditions (Supplementary Table 1). The total number of Dna2 digestion events in different experimental conditions examined over more than three DNA Curtains experiments (n ≥ 3) was indicated in all boxplots. For the boxplot, the red bar represents the median. The minima of the box represents 25^th percentiles, and the maxima is 75^th percentiles. Most extreme data points are covered by the whiskers except outliers. The ‘+’ symbol is used to represent the outliers. Statistical significance was analyzed using the unpaired t-test for two groups. p-value: two-tailed; p-value style: GP: 0.1234 (ns), 0.0332 (*), 0.0021 (**), 0.0002 (***), <0.0001 (****). Confidence level: 95%. f–h Processivity distribution of single DNA2-AC digestion for different experimental conditions (Supplementary Table 1): 130-nm bin. The total number of dna2-AC digestion events (N) was as labeled. Each experimental condition was examined over more than three DNA Curtains experiments (n ≥ 3). Error bars were obtained through the bootstrap analysis. For any normally distributed dataset, 68.27% of the values lie within one standard deviation of the mean, therefore, our choice of 70% confidence intervals for the bootstrapped data provides a close approximation to expectations for one standard deviation from the mean. The data were fitted with Gaussian functions (red dash line). The errors represented 95% confidence intervals obtained through Gaussian function fitting. Source data are provided as a Source Data file.