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. 2018 Oct 25;8:15791. doi: 10.1038/s41598-018-34000-x

Figure 3.

Figure 3

MacExoIII displays the properties conserved in ExoIII family members. DNA substrates were incubated with or without 1 nM MacExoIIIWT or 1 nM MacExoIIIE39A at 37 °C for the indicated times (2, 5, 10, 20 min). Cleavage products were separated by 8 M urea-15% PAGE. M, DNA marker (lanes 1 and 8). (−), no enzyme control (lanes 2 and 9). (A) 5′-Cy5-labeled ssDNA (lanes 1–7) or dsDNA (lanes 8–14). (B) 5′-Cy5-labeled 3′-protruding (lanes 1–7) or 5′-protruding dsDNA (lanes 8–14). (C) 5′-Cy5-labeled (lanes 1–7) or 3′-Cy5-labeled (lanes 8–14) 6-nt 3′-overhang nicked DNA. (D) 5′-Cy5-labeled ssDNA (lanes 1–7) or dsDNA (lanes 8–14) with 3′-phosphate termini. Amounts of cleaved DNA are determined by subtracting amounts of uncleaved substrates (full-length bands) from amounts of the initial substrates on the control lanes, and fractions (%) of cleaved DNA are indicated at the bottom of the gels in italic letters. The cropped gels are used in the figure, and the full-length gels are presented in Supplementary Fig. S9.