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. 2017 May 27;45(12):7226–7236. doi: 10.1093/nar/gkx223

Figure 2.

Figure 2.

Discrete complexes form when AGT binds single-stranded and duplex 13-mer DNAs. Electrophoretic mobility shift assays (EMSAs) were performed at 20 ± 1°C. Panel A: titration of single-stranded 13 nt DNA (2.1 nM) with AGT (ranging from 0 μM to 9.6 μM) to form the low-mobility cooperative complex. The equilibration buffer was 10 mM Tris (pH 7.6 at 20°C), 1 mM DTT, 100 mM KCl, 0.1 mg/ml BSA. Samples were resolved in a 15% native polyacrylamide gel cast and run as described. Band designations: F, free DNA; B, bound DNA. Panel B: Titration of unmodified double stranded 13 bp DNA (1.6 nM) with AGT (ranging from 0 μM to 9.5 μM). Reaction and electrophoresis conditions were as described for Panel A, above. Band designations: F, free DNA; B, bound DNA.