Mutational analysis of human Dicer (hsDicer). (A) Schematic of hsDicer. Domains are shaded. Mutations characterized in this study are indicated, with mutations characterized by small RNA-sequencing underlined. The residue mutated in human nonepithelial ovarian cancers is highlighted in bold. (B) Western blot of Dicer. Differences in signal intensity may reflect differences in immunoreactivity against murine and human Dicer. p107 was used as a loading control. (C) Schematic of the cleavage sites of the RNase IIIA and RNase IIIB domains, and the binding site of the PAZ domain relative to the precursor miRNA hairpin. The placement of the domains is based on findings reported here as well as published findings (Zhang et al. 2004; Park et al. 2011). (D) Northern blot of let-7c. (E) Northern blot of miR-22. The probes and visualized species are indicated to the right of each blot. Migration of a DNA ladder is shown to the left of each blot. tRNAgln was used as a loading control.
