Figure 3. The response of CMG to pre‐formed quadruplexes.

• A
  Pulse‐chase time course experiment with (c‐MYC Pu22)₄ or (SNORD112)₄ substrates. Reactions were initiated with radiolabelled dATP for 10 min, chased with excess ‘cold’ dATP and samples taken at the indicated time points.
• B–E
  CMG unwinding assays on substrates containing a pre‐formed G4 (B), a mutant G4 (see Table EV1) (C), pre‐formed iM (D) or a mutant iM (see Table EV1) (E). CMG unwinding was stimulated by the addition of 2 mM ATP following CMG loading in the presence of ATPγS. Samples were taken at the indicated time points. Products were run on 10% TBE gels. Input and boiled substrates were used as controls to visualise where original and unwound substrates migrate. The proportion of template unwound was calculated by measuring the intensity of the ‘unwound’ product band as a proportion of the total product intensity for each lane.
• F, G
  CMG unwinding assays on duplexed substrates (F) or substrates containing a pre‐formed G4 (G). Reactions were carried out as in (B–E) but with the addition of 0.25 μM PhenDC3 where indicated.

Source data are available online for this figure.