Fig. 4.
Binding studies of Dicer to pre-let-7a-1. (a) Typical EMSA performed with 10 pM of 5′-[32P]-labeled pre-let-7a-1 and increasing concentrations of the D1320A/D1709A Dicer variant (0, 0.1, 0.5, 1, 2.5, 5, 10, 25, 50, 75, 100, 250, 500 and 1000 nM). (b) Typical binding curves of D1320A/D1709A Dicer and WT Dicer to pre-let-7a-1. The data shown here were fitted to the Hill equation to obtain Kd values of 5.1 ± 0.6 nM (WT Dicer) and 8.4 ± 0.4 nM (D1320A/D1709A Dicer) with Hill coefficient of 1.2 ± 0.1 and 1.4 ± 0.1, respectively. Averages from at least three independent experiments yield Kd values of 5 ± 1 nM (WT Dicer) and 9 ± 1 nM (D1320A/D1709A Dicer) with Hill coefficient of 1.3 ± 0.2 and 1.4 ± 0.3, respectively