Fig 2. HopZ3 interacts with members of the PTO defense pathway and PTO activating effectors.
Pull-down experiments using recombinant tagged proteins were performed to assess the interaction between HopZ3 and proteins in the PTO immune pathway. GST, MBP or beads alone were used as negative controls. Proteins were detected by immunoblotting or Coomassie staining. (A) Immobilized GST-tagged FEN and PTO were incubated with purified HopZ3-His, washed and resolved by SDS-PAGE. Immunoblotting showed interactions between these proteins and HopZ3. (B-D) HopZ3 interacts with all three tomato RIN4s. Immobilized MBP-tagged SlRIN4-1 (B) or GST-tagged SlRIN4-2 (C) pulled down HopZ3-His and immobilized HopZ3-GST pulled down SlRIN4-3-His (D). (E) GST-tagged HopZ3 was pulled down using immobilized AvrPto1Psy-His, showing their interaction. (F) HopZ3-GST was pulled down by immobilized AvrPtoBPsy-His. Dashed lines in (A-C) separate input from pull down in the same blot and in (D, F) they mark protein weight standards.