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Fig. 4. T186 is required for interaction with PLK-1PBD 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.