Figure 5.
Interaction assays between WSL4 and OsCER2. A, Isolation of OsCER2 from rice calli. Co-IP performed in transgenic rice calli expressing WSL4-3FLAGs. Plant proteins were separated by SDS-PAGE; the unique band (solid arrow) was excised and analyzed by LC-MS. WSL4-3FLAGs was detected by immunoblotting using an anti-FLAG primary antibody (open arrow). Antitubulin was used as a loading control. Lane 1, wild type; lane 2, expressed WSL4-3FLAGs. B to D, SUY2H assay of WSL4 and OsCER2 with WSL4-, wsl4-1-, and wsl4-2-Cub-LexA-VP16 as bait, and NubG-OsCER2 and NubG-mutated-OsCER2 as prey. Alg5-NubI and Alg5-NubG expression plasmids were used as positive and negative controls, respectively. Yeast transformants were spotted on control medium (–LW) and selective medium (–AHLW; OD yeast cells: 100, 10−1, 10−2, 10−3).