Figure 3
Co-immunoprecipitation of HA-VQ or VQ-RFP proteins and DXS-GFP proteins. (A) In silico prediction of the interaction between DXS and either VQ1 or VQ10 using iFrag algorithm. VQ motifs of VQ1 and VQ10 are represented by yellow boxes. Nicotiana benthamiana leaves from plants were infiltrated with Agrobacterium transformed with (B) 35S:3xHA-VQ1 (HA-VQ1) or 35S:3xHA-VQ10 (HA-VQ10) and 35S:DXS-GFP (DXS-GFP) and (C) 35S:VQ1-RFP(VQ1-RFP) or 35S:VQ10-RFP (VQ10-RFP) and 35S:DXS-GFP (DXS-GFP). By three days after agroinfiltration they were used to make total protein extracts (Input) and to immunoprecipitate (IP) with magnetic beads coated with a-GFP antibodies (B) or magnetic beads coated with a-RFP antibodies (C). Protein extracts were prepared in the presence of 0.5 mM DTT. A high molecular mass oligomer (o) as well as the DXS monomer (m) were detected. Proteins were detected by Western blot with the indicated antibodies. Equal loading was checked in by staining membrane with Ponceau S. The position of molecular mass markers (kDa) is indicated at the left side of each panel.
