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. 2004 Nov 25;24(1):138–148. doi: 10.1038/sj.emboj.7600491

Figure 6.

Figure 6

Analysis of the effects of changes in pri-miR-30a stem length. (A) Predicted hairpin RNA structures adopted by pri-miR-30a variants with different stem extensions beyond the pre-miR-30a structure. Boxed sequences represent added restriction sites. The positions of the 5′ and 3′ ends of the endogenous pre-miR-30a are indicated by arrows in E17. (B) Primer extension experiments to determine the expression level and 5′ ends of mature miR-30a-3p (left panel) or miR-30a-5p (right panel) variants. The sequences of the miR-30a-3p std1 and miR-30a-5p std1 and 2 size standards are given in Supplementary Table 1. (C) The 5′ cleavage sites mapped in panel B are indicated as I, II, III, and IV on the predicted pri-miR-30a RNA structure. Two potential duplex intermediates formed after cleavage are indicated (3′ ends are inferred). (D) In vitro Drosha processing of transcripts encoding E10 and E21. The asterisk indicates the pre-miRNA band. The position of a 66 nt DNA size marker is indicated.