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. 2020 Aug 21;11:1290. doi: 10.3389/fpls.2020.01290

Figure 3.

Figure 3

HopX1i strengthens the ZED1/SZE1 protein-protein interaction. (A) Yeast two-hybrid assay assessing interactions between ZED1 (in pEG202) and PBL15, SZE1, or SZE2 (in pJG4-5) on X-gal reporter plates. To determine protein-protein interaction in the presence of a T3SE, HopX1i, or HopZ1a was integrated into the yeast HO locus (see Methods). (B) Quantitative yeast interaction assays were performed between ZED1 and SZE1 in the presence or absence of the T3SEs HopZ1a (Z1a), HopX1i (X1i), or the catalytic mutant of HopX1i (C198A; X1i C/A). Letters represent statistically significant differences (Tukey’s HSD, P < 0.05). Experiments were replicated three times with similar results. Plasmids or genome integrations expressing each component for every strain used are listed below the bar graph. Strains 5 and 7 were independently generated, but harbor the same combinations of constructs (pBA350V::HopX1i, pEG202::ZED1, and pJG4-5::SZE1). Strains 6 and 8 were independently generated, but harbor the same combinations of constructs (pBA350V::HopX1i C182A, pEG202::ZED1, and pJG4-5::SZE1).