Skip to main content
. 2018 Aug 27;46(17):9057–9066. doi: 10.1093/nar/gky759

Figure 5.

Figure 5.

Stability of SJG-136 crosslinks are DNA length-dependent. Short (A) and long (B) ICL substrates with 3′ fluorescent label. In vitro analysis of SNM1A translesional nuclease activity on short ICL (C) or long ICL (D). SNM1A (0.2 μM) or Lambda exonuclease (5 units) were incubated with duplex DNA substrate (0.2 μM) with (left and middle; lanes 1–8) or without (right; lanes 9–12) an SJG-136 crosslink. Reactions were stopped after 0, 5 and 60 min. Crosslinked products were heat denatured (shown in middle panels) to disrupt the crosslink and allow visualization of the top strand alone. A schematic of products is shown with the dark strand represents the labelled DNA visible on the gel. Products were resolved using 20% denaturing PAGE and imaged with the ChemiDoc XRS (BioRad) at 526 nm. * denotes residual crosslinked DNA following heat treatment. ** denotes unphosphorylated substrate.