Fig. 4. T₁₈₆ is required for interaction with PLK-1^PBD and is important for MEX-5 function. (A) Yeast two-hybrid assays. Duplicated strains were incubated on leucine and tryptophan double dropout medium (Leu- Trp-) either without 3AT (left) or with 50 mM 3AT (right). Each strain tested contains a bait plasmid expressing the protein indicated along the top and a prey plasmid expressing the protein indicated to the left. An unrelated protein, ZYG-11, was used as a negative control for both bait and prey. H/A K/M: H₅₄₂A, K₅₄₄M. (B) Immunofluorescence micrographs of anti-GFP and anti-PIE-1 stainings in mex-5(zu199), or mex-5(zu199);mex-6(RNAi) embryos expressing GFP::MEX-5 T₁₈₆A. Stages indicated to the left. Closed and open arrowheads indicate the germline blastomere and its somatic sister, respectively. (C) Rescue of mex-5(zu199) lethality by GFP::MEX-5 versus GFP::MEX-5 T₁₈₆A, as scored by percentage of viable embryos produced per mex-5 homozygote mother. Scale bar: 10 µm.