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. Author manuscript; available in PMC: 2010 Jul 10.
Published in final edited form as: Cell. 2009 Jul 10;138(1):78–89. doi: 10.1016/j.cell.2009.06.029

FIGURE 3. Characterization of the 5′-flap endonuclease activity of human SLX1-SLX4.

FIGURE 3

A- A 5′ 32P-end labeled stem loop substrate (SL) was incubated for 30 minutes at 30°C with 5 μl S. pombe TEV-Slx1 (Coulon et al., 2004), 150 ng rXPF-ERCC1 or 60 nl FLAG-eluate derived from HEK cells over-producing FLAG-SLX1 and GFP-SLX4. Reaction products were analyzed by denaturing PAGE. The cleavage sites are indicated on the schematized SL.

B- SL was incubated with 60 nl FLAG-eluate (FLAG-SLX1/GFP-SLX4) for 5 min at 30°C in the presence of Mg++ or Mn++.

C- The activity of 30, 60, 120, 240 nl FLAG-eluate derived from HEK cells over-producing FLAG-tagged wild type (WT) or the indicated mutant SLX1 proteins along with GFP-tagged SLX4 was compared on the SL substrate.