Figure 8

Gel mobility shift assay of the recombinant containing APUM5-PHD for 3′ UTR motifs of abiotic stress-responsive genes. (A-D). Indicated ³²P-labeled RNA probes were incubated with recombinant GST-APUM5-PHD (1, 10, 50, and 100 nM) for 30 min in 20 μl RNA binding buffer at room temperature. RNA-protein complexes were separated on a 5% native gel and analyzed by autoradiography. GST protein (1000 nM) was used as a negative control. (E) ³²P-labeled hbNRE2 RNA probes were incubated with recombinant GST-APUM5-PHD (1 and 10 nM) and used as a positive binding control.