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. 2011 May 17;39(16):7161–7178. doi: 10.1093/nar/gkr234

Figure 2.

Figure 2.

Equilibrium binding GMSAs of purified recombinant G4R1/RHAU incubated with unimolecular G4-DNA-containing oligonucleotides yield apparent Kd values in the low picomolar range. (A, B, E, F, I and J) are representative (of three repetitions) phosphorimages of non-denaturing gel electropherograms of GMSAs; the top triangles indicate the direction of increasing G4R1/RHAU concentration. GMSA of 1 pM 5′-[32P]-labeled (A) unimolecular Poly A Zic1 47-mer G4-DNA, (E) unimolecular Poly T Zic1 47-mer G4-DNA, (I) c-Myc 51-mer G4-DNA incubated with a broad range of concentrations of G4R1/RHAU (lane 1, 0 pM; lane 2, 1 pM; lane 3, 5 pM; lane 4, 10 pM; lane 5, 20 pM; lane 6, 40 pM; lane 7, 80 pM). GMSA of 1 pM 5′-[32P]-labeled unimolecular (B) Poly A Zic 47-mer G4-DNA, (F) Poly T Zic1 47-mer G4-DNA, (J) c-Myc 51-mer G4-DNA incubated with concentrations of G4R1/RHAU that were increased in small (0.5–1 pM) increments between (B) lanes 1–12, (F) lanes 1–10, or (J) lanes 1–6, followed by larger increments up to 80 pM. Binding data for Poly A Zic1 47-mer G4-DNA, Poly T Zic1 47-mer G4-DNA, and c-Myc 51-mer G4-DNA were directly fit by non-linear regression to a hyperbolic equation (Panels C, G and K, respectively) and to a double reciprocal linear equation by linear regression (Panels D, H and L, respectively). Error bars are Mean ± SD, n = 3 independent experiments.