FIG 5.

Cdc50 interacts with Crm1. (A) Colocalization of Crm1 and Cdc50. Fluorescent signals generated by Crm1-mCherry and Cdc50-GFP in cells grown in YPD medium. DIC, differential interference contrast. Bar, 5 μm. (B) Interactions between Crm1 and Cdc50 were observed in the split-ubiquitin system. The C-terminal half of ubiquitin (Cub) was fused to the N terminus of Cdc50 cDNA (Cub-Cdc50). The N-terminal half of ubiquitin (NubG) was fused to the N terminus of Crm1 cDNA (NubG-Crm1). Yeast transformants were grown on selective medium. β-Galactosidase activity assays were performed to verify the interaction. Values were averaged from two independent experiments. SD, synthetic dextrose. (C) Total proteins from cells expressing Crm1-mCherry, Cdc50-GFP, or both Crm1-mCherry and Cdc50-GFP were extracted. The potential Crm1-Cdc50 interaction was analyzed by coimmunoprecipitation (IP) with anti-mCherry or anti-GFP antibody and evaluated by immunoblotting.