WormBase Tree Display for Picture: WBPicture0000012578

WBPicture0000012578 Description: Figure S5. Characterization of LIPL-4 Function in Autophagy, Related to Figure 5 (A) Quantification of GFP::LGG-1-positive foci in seam cells of glp-1(e2141) larvae (L3 larva stage) fed control bacteria or bacteria expressing dsRNA against lipl-4 for three 'F2' generations. Cumulative counts of two independent experiments (n 60 seam cells). Data are shown as mean +/- SEM (***P < 0.001 vs. control, t-test). (B) Quantification of GFP::LGG-1 foci in glp-1(e2141) larvae (L3 larva stage) fed control bacteria or bacteria expressing dsRNA for autophagy genes vps-34 or atg-18 for two ('F1') or three ('F2') generations. At each generation analyzed, worms were raised at 25 C from hatching to prevent germline development. Cumulative counts of two independent experiments (n=80-115 seam cells). Data are shown as mean +/- SEM (*P < 0.05, **P < 0.01, ANOVA). (C) Quantification of GFP::LGG-1-positive foci in seam cells of animals overexpressing LIPL-4 (LIPL-4 OE) L3 larvae and in non-transgenic siblings (WT). Animals were raised at 20 C, and mean number of foci +/- SEM is shown (n 280 seam cells, ***P < 0.001, t-test). (D) RT-PCR analysis of bec-1, unc-51, lgg-1, and pha-4 mRNA in 1-day adult WT and LIPL-4 OE animals. Animals were raised at 20 C, and mean expression +/- SD is shown (*P < 0.05, **P < 0.01, ANOVA). (E) Representative picture of a day 1 adult transgenic glp-1(e2141) animals expressing LIPL-4 raised at 25 C. LIPL-4 expression was visualized by GFP co-expression from a polycistronic transcriptional unit (Wang et al., 2008). LIPL-4 expression was seen in the intestinal cells (arrowheads) and hypodermal seam cells (filled arrows), and the expression patterns were similar but of lower magnitude, in glp-1(+) animals (data not shown, see also Supplemental Information). Magnification, 1000-fold. (F) Lifespan analysis of glp-1(e2141) animals fed control bacteria or a 1:1 ratio mix of control bacteria with bacteria expressing vps-34 or lipl-4 dsRNA, or a mix of vps-34 and lipl-4 RNAi bacteria. Experiments were performed twice with similar results; see Table S3 for details and repeats. Animals were raised at 25 C from hatching until the first day of adulthood and were then moved to 20 C for the remainder of their life.
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  Crop: Cropped_from: WBPicture0000012577
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    Anatomy: WBbt:0005753
      WBbt:0005772
  Acknowledgment: Template: Reprinted from <Journal_URL>, <Article_URL>, Copyright <Publication_year>, with permission from <Publisher_URL>.
    Publication_year: 2011
    Article_URL: DOI id 10.1016/j.cub.2011.07.042
    Journal_URL: CurrentBiology
    Publisher_URL: Elsevier
  Reference: WBPaper00040180