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Fig. S2 Birc5b::GFP expression in cultured oocytes and late rescue of motley cytokinesis defects by wild-type Birc5b::GFP mRNA injected post-fertilization. (A–F) Expression of Birc5b::GFP through mRNA injection into in vitro matured oocytes. Immature germinal vesicle containing stage IV oocytes from homozygous motley females express protein from injected birc5b::eGFP mRNA 1 hour post injection (hpi) (A–C). Germinal vesicle dissolution and oocyte clearing occur normally in Birc5b::eGFP-expressing motley oocytes (D–F). (G–K) Partial rescue of cytokinesis defects in motley mutants by expression of Birc5b::GFP through mRNA injection into 1-cell embryos. motley mutant embryos injected with birc5b::eGFP at the 1-cell stage are Birc5b::eGFP-positive at 2 hpf and exhibit several cleavage furrows (G–I) like wild-type embryos (K), although blastomeres in mutants are larger due to the lag in functional rescue through injection at the 1-cell stage. Uninjected sibling motley mutants do not exhibit any furrows (J). (L–O) Rescue of midbody formation defect in motley mutants by expression of Birc5b::GFP through mRNA injection into 1-cell embryos. At 4 hpf, midbodies are seen in wild-type embryos (L, arrows), which are never seen in motley mutants (M). In motley mutants injected with birc5b::eGFP at the 1-cell stage, midbodies are seen by 4 hpf (N, O, arrows). Cells in the panels shown (L–O) are at slightly different stages in the cell cycle (shown by different sizes of asters) due to asynchronicity between embryos and embryonic regions characteristic of these stages [53], [54]. However, midbody structures in the early embryo are stable through multiple cell cycles ([21]; our own observations), allowing a comparison between the various conditions.