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. 2022 Dec 21;42(3):e111998. doi: 10.15252/embj.2022111998

Figure EV3. Comparison of DNA binding and unwinding by WRN and Sgs1 DNA helicases.

Figure EV3

  1. Representative electrophoretic mobility shift assay with WRN and Sgs1, using either pUC19 harboring the random inverted repeat‐based cruciform structure (circular), or linearized pUC19. The samples were run on a 0.8% unstained native agarose gel. The gel was stained after the run with GelRed.
  2. Representative helicase assay with increasing concentrations of WRN and Sgs1, using an oligonucleotide‐based Holliday junction as a substrate. Reactions were supplemented with either human or yeast RPA (15 nM) and analyzed by 10% native acrylamide gel electrophoresis. Red asterisk indicates the position of the radioactive label.
  3. Quantitation of DNA unwinding from assays such as in (B). Averages shown; n ≥ 3; error bars, SEM.
  4. Representative helicase assay with increasing concentrations of WRN and Sgs1, using a 2.2 kbp‐long dsDNA substrate. Reactions were supplemented with either human or yeast RPA and analyzed by electrophoresis on a 1% agarose gel.