Figure 2
In planta Bimolecular Fluorescence Complementation (BiFC) studies on the interaction of RAN1 or type-I ethylene receptors ETR1/ERS1 with soluble copper chaperones ATX1 and CCH. BiFC studies reveal protein interaction of RAN1 and ATX1 (A). Fluorescence complementation was also detected for RAN1 and CCH (B) indicating that both soluble chaperones of the ATX1-family interact with RAN1 in vivo. BiFC studies on the interaction of soluble copper chaperones ATX1 and CCH with type-I ethylene receptor ETR1 and ERS1 indicate a direct protein interaction of receptors with soluble copper chaperones. RFP expression acts as an infiltration control for the BiFC vector and is constitutively expressed (d35S). Constructs containing full-length ETR1 were under control of an inducible promoter, whilst constructs containing ERS1 and ETR11–157 were constitutively expressed (d35S). Arrows indicate the meshed like structure or nucleus envelope staining, typical for the ER, visible only for the complementation signal (YFP) but not for free RFP. nYFP and cYFP symbolize the YFP fragment fused either N- or C-terminal to the protein of interest.
