Fig. 1

AvrPik-D interacts with WG7. A Analysis of protein–protein interactions between BD:AvrPik-D (ΔSP) and AD:WG7 via yeast-two-hybrid (Y2H) assay. Yeast cells harboring the indicated bait and prey plasmids were diluted 1 × , 10 × , and 100 × , and then spotted onto selective media SD-Leu-Trp and SD-Leu-Trp-His-Ade with X-α-gal. The positive and negative controls were pGADT7-T + pGBKT7-53 and pGADT7-T + pGBKT7-lam. SD (-Leu-Trp) indicates selective medium lacking leucine and tryptophan. SD (-Leu-Trp-His-Ade) indicates selective medium lacking leucine, tryptophan, adenine and histidine. B Deletion assays of WG7 to test interactions with AvrPik-D. A series of truncated WG7 proteins were tested for interaction with AvrPik-D via Y2H. C In vitro pull-down assays were conducted with fusion proteins (GST:WG7¹⁻⁶⁹⁸ and MBP:AvrPik-D) using GST beads and detected by immunoblot analysis with anti-GST and anti-MBP antibodies. D Co-immunoprecipitation (Co-IP) assay of WG7N-GFP and AvrPik-D-mCherry in N. benthamiana. The Co-IP assay (IP) was carried out with anti-GFP beads, and the proteins were analyzed by western blot with anti-GFP and anti-mCherry antibodies. E Y2H assay to detect the interaction among WG7¹⁻¹⁶⁰⁰, WG7¹⁻⁶⁹⁸, WG7^699−1600, Pikh1 and Pikh2.