FIGURE 5.

Pulldown assays allow identification of the OsNug2 domain that interacts with OsL10a. A, schematic representation of OsNug2 deletion variants. B, pulldown assay of GST-OsNug2 with [³⁵S]methionine-labeled OsL10a. Radiolabeled OsL10a was incubated with GST alone (lane 2) or GST-OsNug2 (lane 3). Reaction mixtures were incubated with glutathione beads, centrifuged, and the supernatants were discarded. Residual precipitates were boiled and subjected to SDS-PAGE. Five percent of total ³⁵S-labeled OsL10a was loaded into lane 1 and 50% was used for the pulldown assay (lane 3). C, pulldown assay of [³⁵S]methionine-labeled OsNug2 deletion mutants with GST-OsL10a. Ten percent of total ³⁵S-labeled OsNug2 variants were loaded into lane 1 and 50% was used for the pulldown assay (lane 3).