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WBPicture0000014754 Description: Figure 1. alfa-1 Mutations Cause a Defect in Yolk Storage or Transport in the C. elegans Embryo (A) Genomic organization of alfa-1. Two alleles of alfa-1, gk498021 and ok3062, are both likely to be null. gk498021 is a G-to-A transition leading to a tryptophanto-opal stop codon at amino acid 54, and ok3062 is a partial deletion of exons 3 and 4. Black bar, sequence deleted in the ok3062 mutant.(B) Differential interference contrast (DIC) and confocal images of wild-type, alfa-1(ok3062), and alfa-1(gk498021) comma-stage (top) and 3-fold (bottom) embryos containing the yolk protein transgene pwIs98[vit-2::tdimer2] or nIs755[vit-2::bfp, vit-2::pHluorin], respectively. Arrowheads, blobs (abnormal yolk presencein extra-embryonic fluid). Scale bars, 10 um.(C) Confocal images of comma-stage embryos of the indicated genotypes containing the transgene nIs755[vit-2::bfp, vit-2::pHluorin]. Mutation of rme-2, theoocyte's yolk receptor gene, suppresses yolk uptake and the blob phenotype. Scale bar, 10 um. Animals also contained the reporter qxIs257[nuc-1::mCherry]. (D) Percentage of 3-fold or older embryos of the indicated genotypes containing blobs, showing the suppression of the alfa-1 blob phenotype caused by loss ofthe yolk receptor gene rme-2 function. For the quantification of the penetrances of the blob phenotype, we studied embryos at the 3-fold stage or older but beforethe initiation of pumping and derived from 1-day adults, unless otherwise stated. Nomarski differential interference contrast (DIC) optics was used.(E) Survival curve of wild-type (black), alfa-1(ok3062) (red), and alfa-1(gk498021) (blue) L1 larvae that hatched in the absence of food. The data shown arerepresentative of two independent experiments.(F) Anti-HA tag immunofluorescence of proliferative (left) and comma-stage (right) embryos expressing the reporter nIs759[Palfa-1::alfa-1 gDNA::HA::alfa-1 3'UTR]showing ALFA-1::HA cytoplasmic localization. Scale bar, 10 mm.(G) Percentage of 3-fold or older embryos of the indicated genotypes containing blobs. Overexpression of either wild-type alfa-1 or codon-optimized C9orf72 forexpression in C. elegans (ceC9orf72) completely or partially rescued the alfa-1 blob phenotype, respectively. z, animals expressed a Palfa-1::4xnls::gfp::alfa-130UTR transcriptional reporter. See also Figure S1.
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  Crop: Crop_picture: WBPicture0000014755
  Acknowledgment: Template: Reprinted from <Journal_URL>, <Article_URL>, Copyright <Publication_year>, with permission from <Publisher_URL>.
    Publication_year: 2018
    Article_URL: DOI id 10.1016/j.cub.2018.03.063
    Journal_URL: CurrentBiology
    Publisher_URL: Elsevier
  Reference: WBPaper00054449