Fig. 2. Dicer interacts with dsRNA in EMSA. (A) ³²P-labeled dsRNA was incubated in the absence or presence of Dicer (no MgCl₂ added), (B) without or with unlabeled dsRNA or dsDNA poly(dA·dT), or (C) without or with anti-Dicer antibody (0.5–4.0 µl; no MgCl₂ added). (D) ³²P- labeled CLP 23 nt siRNA or 5LO dsRNA (100 bp) was incubated in the absence or presence of Dicer. (E) Left panel, ³²P-labeled dsRNA was incubated in the absence or presence of Dicer, at the indicated temperatures for 1 h prior to EMSA analysis. Middle panel, same as left panel, but run half way. Dicer·dsRNA complex formation was analyzed by non-denaturing PAGE and autoradiography. Right panel, RNA was extracted from the indicated bands and analyzed by denaturing PAGE and autoradiography. The 20–30 nt region is shown, with the full-length gel on the right.