Figure 2.

Gel Mobility Shift Assays for Detection of sRNA Binding Affinity of SD2b and Its Mutants.

sRNA binding affinity assays. GST-tagged SD2b ([A] and [B]), NLS mutants ([E] and [F]), and deletion mutants ([C] to [F]) were incubated with [γ-32P]-labeled synthetic 21- and 24-nucleotide siRNA ([A], [C], and [D]), miRNA duplexes (B), or with [γ-32P]-labeled synthetic 55-nucleotide-long ss or ds RNA ([E] and [F]) as indicated above the gels. GST-tagged P19 (P19), GST (G), and buffer (-) were used as controls. One nanomole of purified proteins was loaded in each assay. Bound and free probes are indicated. nt, nucleotides.