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. Author manuscript; available in PMC: 2021 Nov 16.
Published in final edited form as: Chem Res Toxicol. 2020 Nov 5;33(11):2903–2913. doi: 10.1021/acs.chemrestox.0c00413

Figure 1.

Figure 1.

Unwinding DNA and release of a reversible interstrand cross-link. (A) A replication fork model (5′-[32P]-OD1:OD2, 10 nM) containing an interstrand cross-link formed by bisQMP and a radiolabel on the translocated strand was incubated in the alternative and combined presence of T7gp4 (55 nM monomer) and dTTP (1 mM). (B) After incubation at 37 °C for the indicated time, reaction was quenched by addition of EDTA (40 mM) and analyzed by denaturing polyacrylamide gel electrophoresis (PAGE, 10%). As electrophoretic standards, lane ss contains only 5′-[32P]-OD1 and lane xl contains 5′-[32P]-OD1:OD2 cross-linked by bisQMP. (C) The remaining cross-link (% based on total radiolabel) was determined by phosphoimagery in three replicates and their average values are indicated by the cross-bars.