Processing of pre-mmu-mir-1982 RNA by recombinant DICER protein. (A) Pre-mmu-mir-1982 RNA and probes used in this study. The secondary structure was predicted using the CentroidFold program [49]. The solid line shows the position of probe-4 and the dashed line shows the position of probe-5. (B-C) Time-course analysis of the processing of pre-mmu-mir-1982 RNA by the rDICER protein. pre-mmu-mir-1982 RNAs were incubated with rDICER in vitro for the indicated time points (0, 10, 20, 30, 40, 50, 60, 75, 90, 105 and 120 min). The RNAs processed by rDICER were detected using probe-4 (B), probe-5 (C) by Northern blotting. The gray arrow shows the band of unprocessed RNA and the black arrow shows the bands of small RNA processed from the 5' strand and 3' strand of pre-mmu-mir-1982 RNA respectively. M: decade marker.