Figure 8.

CES is a nuclear protein that interacts with BEE1 and is phosphorylated by BIN2 in vitro. (A) CES–YFP reporter expression in Arabidopsis protoplasts treated with 24-epiBL (1 μM) or Bkn (30 μM) for 2 h as compared with an untreated control. (B) Colocalization of CES–CFP and BEE1–YFP. Images of a representative protoplast coexpressing 35S_p:CES-CFP and 35S_p:BEE1-YFP, treated for 2 h with 1 μM of 24-epiBL. (C) Bimolecular fluorescence complementation assay showing a representative protoplast cotransformed with CES–nYFP and BEE1–cYFP constructs. (D) In vitro kinase assays using 0.1 μg of GST–BIN2 and 1.0 μg of GST–CES. The reactions were treated for 2 h with increasing concentrations of Bkn. A reaction to which only BIN2 was added served as a negative control.