Figure 5.

In vitro processing of 5′-extended pre-miRNAs by human Dicer. (A) Equal numbers of in vitro transcribed 5′-extended pre-miR-HSUR4s were incubated with wild type (WT) or transdominant negative (TN) human Dicer and analyzed by northern blot to detect miR-HSUR4-5p or -3p. The asterisk (*) marks the 22 nt miR-HSUR4-5p resulting from secondary Dicer cleavage (see Supplementary Figure S9). (B) Quantitations of miR-HSUR4-5p/-3p ratio from Figures 2B and 5A (mean ± standard deviation) were derived from three independent experiments. In vitro miR-HSUR4-5p/-3p ratio (▪) were derived from Figure 5A. In vivo miR-HSUR4-5p/-3p ratio (□) were derived from Figure 2B. ND: not determined due to insufficient signal. (C) Quantitations of relative mature miRNA levels (mature miRNA/pre-miRNA) compared to WT pre-miR-HSUR4 in (A) (mean ± standard deviation) were derived from three independent experiments. FC: fold change.