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Figure S2. Characterization of sterile phenotype of the spdl-1 deletion strain and cell cycle–dependent alteration of the subcellular localization of SPDL-1 in one-cell–stage embryos. (A) Single gonad arms (outlined in red) of an adult hermaphrodite homozygous for spdl-1(ok1515) (spdl-1Δ) and an N2 adult hermaphrodite were dissected, fixed with methanol and acetone, and stained with 100 ng/ml DAPI. The distal end of each gonad is indicated by a red asterisk. The endomitotic oocytes observed in the spdl-1Δ gonad are indicated with red arrows. (B) Chromosomes in germ cells at diakinesis in spdl-1Δ homozygotes and N2 worms were stained with DAPI. The number of bivalents in the nuclei is shown. (C) The bivalents in 30 nuclei of spdl-1Δ germ cells and 60 nuclei of N2 germ cells were analyzed. The number of nuclei that have six bivalents (normal) and more than six bivalents (aneuploid) are shown. (D) The sterile phenotype of spdl-1Δ homozygotes was suppressed by the temperature-sensitive allele of emb-30, emb-30(tn377) (emb-30ts). L4 young adult hermaphrodites with indicated genotypes segregated from heterozygous parents were individually plated and incubated at 20°C for 12 h, and then the percentage of worms that laid eggs was calculated. (E) Whole-worm extracts were prepared from wild-type (WT) and spdl-1 deletion (spdl-1∆) strains and subjected to Western blot, using an affinity-purified anti–SPDL-1 antibody. SPDL-1 antibody specifically identifies cellular SPDL-1 in whole-worm lysates. (F) Immunofluorescence images of fixed embryos at indicated stages of the first mitosis. DNA (white), tubulin (green), and SPDL-1 (red) were stained with DAPI, antitubulin, and anti–SPDL-1 antibodies, respectively. SPDL-1 colocalizes to microtubules through mitosis. SPDL-1 also temporarily localizes to kinetochores from prometaphase until anaphase. Bars, 20 µm.