Figure 2

Recombinant Ago2 and Dicer are sufficient to recapitulate processing of 5'P -pre-miR- 24-1 and loading of miR-24-1* strand into Ago2 in vitro. A. Results of cleavage assays using recombinant Ago2, Dicer, and TRBP (in combinations as indicated) pre-incubated with 5' -P-pre-miR-24-1, prior to the addition of radiolabeled 3' -arm target (Ta-3). B. Experiments were performed as in (A), except with radiolabeled pre-miR-24-1 and unlabeled 3' -arm target (Ta-3). 50% of the total volume of each reaction was analyzed on 15% Urea PAGE and RNAs were detected by autoradiography. C. The remaining 50% of each reaction was analyzed by native RNA gel electrophoresis. Pre-annealed 22nt RNA duplex was used as size marker.