Figure 7.

PLDγ1 can be found in complex with epitope-tagged BIR2 and BIR3. Western blot analysis of transiently expressed proteins in N. benthamiana 3 d after infiltration. Leaf material was harvested 5 min after treatment with 1 µm flg22 (+) or water (−). After protein extraction, the proteins were subjected to immunoprecipitation (IP) with GFP- or myc-affinity beads as indicated. For different immunoprecipitations within one experiment (A and D), the same source material was used. Immunoprecipitated and copurified proteins were detected with tag-specific antibodies as indicated. A and D, Coimmunoprecipitation of BIR2-myc (A) or BIR3-myc (D) and PLDγ1-GFP using myc-trap beads or GFP-trap beads, respectively. B and E, Copurification of BIR2-GFP (B) and BIR3-GFP (E) and PLDγ1-myc precipitated with GFP-trap beads. C and F, Copurification of BIR2-GFP (C) and BIR3-GFP (F) and PLDγ1-HA precipitated with GFP-trap beads. Numbers in the images indicate quantification of signal intensities. All experiments were repeated at least three times with similar results.