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. 2014 Oct 10;5:520. doi: 10.3389/fpls.2014.00520

Figure 4.

Figure 4

Interaction analyses among S(ΔH) K3, OpsDHN1 (SK3), and truncated versions (SK2 and S) using the split-ubiquitin yeast two-hybrid assay. (A) Schematic representations of the OpsDHN1 S(ΔH)SK3 version without histidine-rich motif (residues 1–111:128–248). (B) Yeast cells expressing the control system interaction LargeT-Cub/Δp53-NubG, OpsDHN1 SK3 full-length interaction. Interaction analysis among bait OpsDHN1 S(ΔH) K3-Cub with OpsDHN1-NubG [SK3, S(ΔH) K3, SK2, and S] prey vectors, interaction analysis of the swapped versions in the respective bait and prey vectors. Yeast strains was plated to an OD600 of 0.8, and at serial 10-fold dilutions on semi-selective (SD-LW) and on selective (SD-LWHA) media supplemented with 3-AT (45 and 55 mM). Quantitative β-Galactosidase activity was assayed by hydrolysis of the o-nitrophenyl-b-galactoside (ONPG), as expressed in nmol ONP/min per mg of protein. Data represent the mean ± SD, (n = 3).