Figure 4.

VOZ1 and VOZ2 interact with CO in vitro. A and B, GST pull-down assay showing the interaction of VOZ1 and VOZ2 with CO. His-VOZ1 (A) or His-VOZ2 (B) proteins were incubated with GST or GST-CO together with glutathione-Sepharose beads in a column. The bead-bound proteins were subjected to 10% (w/v) SDS-PAGE and detected by immunoblot (IB) analysis using anti-His antibody. For the input blots, 0.5% (v/v) input extracts were loaded to detect His-VOZ1 or His-VOZ2. C, Electrophoretic mobility shift assay (EMSA) showing the interaction of VOZ1 and VOZ2 with CO. [γ-³²P]ATP-labeled wild-type (WT) probe containing the VOZ2-binding site (GCGTGTGATACACGT) was incubated with His-VOZ2 alone or in combination with GST-CO or GST. As controls, GST-CO and GST alone were incubated separately with the wild-type probe. Additionally, His-VOZ2 was incubated with a probe containing a mutant VOZ2-binding site (tCGTGTGATACACGT).