Somatic aging pathways regulate reproductive plasticity in Caenorhabditis elegans
- Department of Biology, Syracuse University, United States
Figures
Figure 1 with 1 supplement
Adult reproductive plasticity is dependent on DAF-12 steroid signaling.
(A) Model of DAF-12 regulation of development. See text for details. (B) Brood size of CON and PDStv in wild-type N2 and mutant strains. * p < 0.05, ** p < 0.01, and **** p < 0.0001 compare CON and …
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Figure 1—source data 1
Dafachronic acid-dependent DAF-12 signaling is required for decreased fecundity after starvation-induced dauer formation.
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig1-data1-v2.xlsx
Figure 1—figure supplement 1
Dafachronic acid affects adult reproductive plasticity.
(A) Brood size comparison between PDStv grown in the presence of Δ7-dafachronic acid (Δ7-DA) (PDStv + DA) and PDStv grown in the absence of Δ7-DA (PDStv - DA) in wild-type N2, daf-9(rh50), daf-36(k11…
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Figure 1—figure supplement 1—source data 1
Brood size comparison between PDStv grown in the presence of delta7-dafachronic acid (PDStv + DA) and PDStv grown in the absence of (delta7-DA (PDStv - DA).
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig1-figsupp1-data1-v2.xlsx
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Figure 1—figure supplement 1—source data 2
Brood size comparison of PDStv relative to CON.
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig1-figsupp1-data2-v2.xlsx
Figure 2 with 2 supplements
TCER-1 and KRI-1 regulate the decreased fecundity phenotype in PDStv adults.
(A) Model of the regulation of DAF-16 nuclear localization. See text for details. (B) Brood size of CON and PDStv in wild-type N2, daf-18(e1375), pqm-1(ok485), daf-16(mu86); daf-16AM::gfp, smk-1(mn15…
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Figure 2—source data 1
TCER-1 and KRI-1 regulate the decreased fecundity phenotype in PDStv adults.
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig2-data1-v2.xlsx
Figure 2—figure supplement 1
Differentially expressed genes in wild-type N2 PDStv are enriched for DAF-16 targets.
(A) Comparison of DAF-16 and PQM-1 targets (Tepper et al., 2013) with differentially expressed genes in N2 PDStv to N2 CON (Ow et al., 2018). See Figure 2—figure supplement 1—source data 1. (B) …
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Figure 2—figure supplement 1—source data 1
Comparison of DAF-16 and PQM-1 targets (Tepper et al., 2013) with differentially expressed genes in N2 PDStv to N2 CON (Ow et al., 2018).
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig2-figsupp1-data1-v2.xlsx
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Figure 2—figure supplement 1—source data 2
Comparison of DAF-16 and TCER-1 targets with functions in lipid metabolism from germ line-less glp-1(e2141ts) animals (Amrit et al., 2016) with differentially expressed genes in N2 PDStv to N2 CON (Ow et al., 2018).
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig2-figsupp1-data2-v2.xlsx
Figure 2—figure supplement 2
Localization of DAF-16::GFP in CON and PDStv one-day-old adults in (A) CF1139 (daf-16(mu86) I; muIs61 [(pKL78) daf-16::gfp + rol-6(su1006)]) and (B) TJ356 (zIs356 [daf-16p::daf-16a/b::gfp + rol-6(su1006)]).
The GPF observed in CF1139 CON is intestinal background signal.
Figure 3 with 1 supplement
Fatty acid metabolism pathways modulate adult reproductive plasticity.
(A, B, C) Brood sizes of CON and PDStv in wild-type N2 and mutant strains. *** p < 0.001 and ****p < 0.0001 compare CON and PDStv within a genotype; &&p < 0.01 and &&&& p < 0.0001 compare N2 CON to …
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Figure 3—source data 1
Fatty acid metabolism pathways modulate adult reproductive plasticity.
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig3-data1-v2.xlsx
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Figure 3—source data 2
Fatty acid metabolism pathways modulate adult reproductive plasticity.
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig3-data2-v2.xlsx
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Figure 3—source data 3
Fatty acid metabolism pathways modulate adult reproductive plasticity.
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig3-data3-v2.xlsx
Figure 3—figure supplement 1
Fatty acid metabolism in C. elegans.
(A) Model of oleic acid synthesis. NHRs refer to NHR-80, NHR-13, and NHR-66. (B) Model of fat metabolism. Saturated fatty acids, SFAs (blue): C16:0 (palmitic), C18:0 (stearic); monounsaturated fatty …
Figure 4 with 1 supplement
Lipid metabolism is affected in wild-type postdauer adults that experienced starvation-induced dauer.
(A) Brood size of wild-type N2 PDStv and mutant PDStv with or without oleic acid (OA) supplementation. * p < 0.05 and **** p < 0.0001 compare PDStv to PDStv + OA within a genotype; & p < 0.05, &&p < …
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Figure 4—source data 1
Brood size of wild-type N2 PDStv and mutant PDStv with or without oleic acid (OA) supplementation.
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig4-data1-v2.xlsx
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Figure 4—source data 2
Oil Red O (ORO) intensity in CON and PDStv one-day-old adults.
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig4-data2-v2.xlsx
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Figure 4—source data 3
ORO intensity of embryos measured in utero in one-day-adults.
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig4-data3-v2.xlsx
Figure 4—figure supplement 1
Postdauer one-day-old wild-type N2 adults exhibit fat-stained embryos despite the absence of intestinal fat stores.
(A) Representative micrographs of ORO-stained wild-type N2 CON and PDStv. Scale bar: 50 μM. (B) Representative micrographs of ORO-stained wild-type N2, daf-12(rh284), daf-12(rh285), and tcer-1(tm1452…
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Figure 4—figure supplement 1—source data 1
Fatty acid composition of N2 CON and PDStv one-day-old adults.
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig4-figsupp1-data1-v2.xlsx
Figure 5 with 3 supplements
Vitellogenesis and adult lifespan are affected in postdauer animals.
(A) ORO staining in N2 CON and PDStv one-day-old adults fed with vit-1 RNAi or control empty RNAi vector (EV). **** p < 0.0001 compares CON and PDStv of the same RNAi condition; &&p < 0.01 and ####p …
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Figure 5—source data 1
ORO staining in N2 CON and PDStv one-day-adults fed with vit-1 RNAi or control empty RNAi vector (EV).
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig5-data1-v2.xlsx
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Figure 5—source data 2
Adult lifespan assay of N2 CON and PDStv P0 and F1 generations.
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig5-data2-v2.xlsx
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Figure 5—source data 3
Adult lifespan assay of N2 CON and PDStv fed with vit-1 or control empty vector RNAi.
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig5-data3-v2.xlsx
Figure 5—figure supplement 1
mRNA level of vit-1, vit-2, vit-3/4/5, and vit-6 in one-day-old N2 CON and PDStv adults as measured by reverse transcription quantitative PCR (RT-qPCR) following RNAi-by-feeding with k09f5.2 (vit-1) or control empty RNAi vector.
act-1 was used as the normalization control. **** p < 0.0001 and ns (no significance) refer to the comparison of PDStv to CON for a given RNAi knockdown; two-way ANOVA with Sidak’s multiple …
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Figure 5—figure supplement 1—source data 1
mRNA level of vit-1, vit-2, vit-3/4/5, and vit-6 in one-day-old N2 CON and PDStv adults following RNAi-by-feeding with vit-1 or control empty RNAi vector.
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig5-figsupp1-data1-v2.xlsx
Figure 5—figure supplement 2
Altered vitellogenesis does not affect fecundity in postdauer animals.
Brood size of N2 CON and PDStv fed with vit-1 or control empty vector (EV) RNAi. ****p < 0.0001 compares CON and PDStv of the same RNAi condition; and p < 0.05 compares N2 CON vit-1 RNAi to CON EV …
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Figure 5—figure supplement 2—source data 1
Altered vitellogenesis does not affect fecundity in postdauer animals.
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig5-figsupp2-data1-v2.xlsx
Figure 5—figure supplement 3
Reproductive plasticity in postdauer animals is not subject to transgenerational inheritance.
(A) Transgenerational brood size (P0, F1, and F2 generations) of wild-type N2 CON and PDStv. ****p < 0.0001 compares CON and PDStv of the same generation; &&&&p < 0.0001 compares the P0 with the F1, …
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Figure 5—figure supplement 3—source data 1
Reproductive plasticity in postdauer animals is not subject to transgenerational inheritance.
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig5-figsupp3-data1-v2.xlsx
Figure 6 with 1 supplement
Generational inheritance of starvation memory is dependent upon germline-specific RNAi pathways.
(A) ORO staining in wild-type N2, hrde-1(tm1200), and prg-1(tm872) CON and PDStv one-day-old adults from P0, F1, and F2 generations. *p < 0.05 and ****p < 0.0001 compares CON (C) and PDStv (P) …
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Figure 6—source data 1
ORO staining in wild-type N2, hrde-1(tm1200), and prg-1(tm872) CON and PDStv one-day-old adults from P0, F1, and F2 generations.
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig6-data1-v2.xlsx
Figure 6—figure supplement 1
Representative micrographs of N2, hrde-1(tm1200), and prg-1(tm872) CON and PDStv from P0, F1, and F2 generations stained with ORO and quantified in Figure 6A.
Scale bar: 250 μM.
Figure 7 with 2 supplements
DAF-36 and TCER-1 regulate the onset of germline proliferation.
Total germ cell rows in CON and PDStv wild-type N2 and mutant larva exhibiting L3 vulva morphology (see Materials and Methods). * p < 0.05 and **** p < 0.0001 compare CON and PDStv within a genotype;…
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Figure 7—source data 1
Total germ cell rows in CON and PDStv wild-type N2 and mutant larva.
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig7-data1-v2.xlsx
Figure 7—figure supplement 1
DAF-36 and TCER-1 are required for the delay in the onset of germline proliferation in postdauer animals.
(A) Mitotic cell rows at the L3 stage in CON and PDStv wild-type N2, daf-36(k114), nhr-49(nr2041), and tcer-1(tm1452) was determined by subtracting the meiotic cell rows from the total cell rows for …
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Figure 7—figure supplement 1—source data 1
DAF-36 and TCER-1 are required for the delay in the onset of germline proliferation in postdauer animals.
- https://cdn.elifesciences.org/articles/61459/elife-61459-fig7-figsupp1-data1-v2.xlsx
Figure 7—figure supplement 2
Model of endocrine signaling pathways acting as contributors to fatty acid metabolism of PDStv animals.
Signaling between the soma and the germ line establishing metabolic plasticity and the transmission of ancestral starvation memory are dependent on the germline nuclear RNAi pathway.
Appendix 1—figure 1
GLP-4 is required in multiple tissues to regulate adult reproductive plasticity.
Brood size comparison of PDStv relative to CON in wild-type N2, glp-4(bn2), and tissue-specific MosSCI rescue strains of glp-4. Assays were done at 15°C. ** p < 0.01, *** p < 0.001, and **** p < …
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Appendix 1—figure 1—source data 1
GLP-4 is required in multiple tissues to regulate adult reproductive plasticity.
- https://cdn.elifesciences.org/articles/61459/elife-61459-app1-fig1-data1-v2.xlsx
Appendix 1—figure 2
KRI-1 regulates the onset of germline proliferation.
Total, mitotic, and meiotic cell rows in control and PDStv animals exhibiting L3 vulva morphology in wild-type N2 and kri-1(ok1251). **** p < 0.0001 compares of CON and PDStv within a genotype; &&&&p…
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Appendix 1—figure 2—source data 1
Total, mitotic, and meiotic cell rows in control and PDStv animals in wild type and kri-1(ok1254).
- https://cdn.elifesciences.org/articles/61459/elife-61459-app1-fig2-data1-v2.xlsx
Appendix 1—figure 3
Decreased fecundity in postdauer animals results from a reduction of sperm available for self-fertilization.
Brood size of wild-type N2 CON and PDStv self-fertilized hermaphrodites (⚥) and CON or PDStv hermaphrodites mated with CON males (♂). **** p < 0.0001 compares self-fertilized CON or PDStv …
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Appendix 1—figure 3—source data 1
Decreased fecundity in postdauer animals results from a reduction of sperm available for self-fertilization.
- https://cdn.elifesciences.org/articles/61459/elife-61459-app1-fig3-data1-v2.xlsx
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (include species here) | ||||
| Strain, strain background (Caenorhabditis elegans) | N2 | Caenorhabditis Genetics Center | Wild type | |
| Strain, strain background (Caenorhabditis elegans) | AA82 | Caenorhabditis Genetics Center | daf-12(rh284) X | |
| Strain, strain background (Caenorhabditis elegans) | AA85 | Caenorhabditis Genetics Center | daf-12(rh285) X | |
| Strain, strain background (Caenorhabditis elegans) | AA292 | Caenorhabditis Genetics Center | daf-36(k114) V | |
| Strain, strain background (Caenorhabditis elegans) | AA1052 | Adam Antebi, Max Planck Institute | dhs-16(tm1890) V | |
| Strain, strain background (Caenorhabditis elegans) | AE501 | Caenorhabditis Genetics Center | nhr-8(ok186) IV | |
| Strain, strain background (Caenorhabditis elegans) | BS1080 | Tim Schedl, Washington University | gld-1::gfp::3xflag | |
| Strain, strain background (Caenorhabditis elegans) | BX26 | Caenorhabditis Genetics Center | fat-2(wa17) IV | |
| Strain, strain background (Caenorhabditis elegans) | BX106 | Caenorhabditis Genetics Center | fat-6(tm331) IV | |
| Strain, strain background (Caenorhabditis elegans) | BX107 | Caenorhabditis Genetics Center | fat-5(tm420) V | |
| Strain, strain background (Caenorhabditis elegans) | BX110 | Caenorhabditis Genetics Center | fat-6(tm331) IV; fat-5(tm420) V | |
| Strain, strain background (Caenorhabditis elegans) | BX153 | Caenorhabditis Genetics Center | fat-7(wa36) V | |
| Strain, strain background (Caenorhabditis elegans) | BX156 | Caenorhabditis Genetics Center | fat-6(tm331) IV; fat-7(wa36) V | |
| Strain, strain background (Caenorhabditis elegans) | BX160 | Caenorhabditis Genetics Center | fat-7(wa36) fat-5(tm420) V | |
| Strain, strain background (Caenorhabditis elegans) | CB1375 | Caenorhabditis Genetics Center | daf-18(e1375) IV | |
| Strain, strain background (Caenorhabditis elegans) | CE541 | Caenorhabditis Genetics Center | sbp-1(ep79) III | |
| Strain, strain background (Caenorhabditis elegans) | CF1139 | Caenorhabditis Genetics Center | daf-16(mu86) I; muIs61 [(pKL78) daf16::gfp + rol-6(su1006)] | |
| Strain, strain background (Caenorhabditis elegans) | CF2052 | Caenorhabditis Genetics Center | kri-1(ok1251) I | |
| Strain, strain background (Caenorhabditis elegans) | CF2167 | Caenorhabditis Genetics Center | tcer-1(tm1452) II | |
| Strain, strain background (Caenorhabditis elegans) | EG6699 | Caenorhabditis Genetics Center | ttTi5605 II; unc-119(ed3) III; oxEx1578 [eft-3p::gfp + Cbr-unc-119] | |
| Strain, strain background (Caenorhabditis elegans) | GR2063 | Caenorhabditis Genetics Center | hsd-1(mg433) I | |
| Strain, strain background (Caenorhabditis elegans) | RG1228 | Caenorhabditis Genetics Center | daf-9(rh50) X | |
| Strain, strain background (Caenorhabditis elegans) | SEH301 | This study | nhr-13(gk796) V backcrossed | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH302 | This study | nhr-49(nr2041) I; nhr-80(tm1011) III | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH303 | This study | nhr-49(nr2041) I; nhr-13(gk796) V | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH304 | This study | nhr-49(nr2041) I; nhr-80(tm1011) III; nhr-13(gk796) V | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH312 | This study | daf-16(mu86) I; muEx158 (daf-16AM::gfp + sur-5p::gfp) | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH319 | This study | nhr-49(et8) I backcrossed | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH326 | This study | nhr-49(et13) I backcrossed | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH327 | This study | nhr-49(et7) I backcrossed | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH342 | This study | nhr-49(nr2041) I; nhr-66(ok940) IV | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH343 | This study | nhr-49(gk405) I backcrossed | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH344 | This study | nhr-49(ok2165) I backcrossed | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH350 | This study | pqm-1(ok485) II backcrossed | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH351 | This study | kri-1(ok1251) I; daf-12(rh284) X | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH352 | This study | kri-1(ok1251) I; daf-12(rh285) X | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH353 | This study | tcer-1(tm1452) II; daf-12(rh284) X | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH354 | This study | tcer-1(tm1452) II; daf-12(rh285) X | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH357 | This study | glp-4(bn2) I; pdrSi1 [Pglp-4::glp-4 cDNA::gfp::glp-4 3'UTR; unc-119(+)] II | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH368 | This study | glp-4(bn2) I; pdrSi2 [Pnhx-2::glp-4 cDNA::gfp::glp-4 3'UTR; unc-119(+)] II | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH369 | This study | glp-4(bn2) I; pdrSi3 [Pzfp-2::glp-4 cDNA::gfp::glp-4 3'UTR; unc-119(+)] II | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH370 | This study | glp-4(bn2) I; pdrSi4 [Ppgl-1::glp-4 cDNA::gfp::glp-4 3'UTR; unc-119(+)] II | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SEH383 | This study | hrde-1(tm1200) III backcrossed | Sarah Hall, Syracuse University |
| Strain, strain background (Caenorhabditis elegans) | SS104 | Caenorhabditis Genetics Center | glp-4(bn2) I | |
| Strain, strain background (Caenorhabditis elegans) | SP488 | Caenorhabditis Genetics Center | smk-1(mn156) V | |
| Strain, strain background (Caenorhabditis elegans) | STE68 | Caenorhabditis Genetics Center | nhr-49(nr2041) I | |
| Strain, strain background (Caenorhabditis elegans) | STE69 | Caenorhabditis Genetics Center | nhr-66(ok940) IV | |
| Strain, strain background (Caenorhabditis elegans) | STE70 | Caenorhabditis Genetics Center | nhr-80(tm1011) III | |
| Strain, strain background (Caenorhabditis elegans) | STE73 | Caenorhabditis Genetics Center | nhr-80(tm1011) III; nhr-13(gk796) V | |
| Strain, strain background (Caenorhabditis elegans) | TJ356 | Caenorhabditis Genetics Center | zIs356 [daf-16p::daf-16a/b::gfp + rol-6(su1006)] | |
| Strain, strain background (Caenorhabditis elegans) | WM161 | Caenorhabditis Genetics Center | prg-1(tm872) II | |
| Strain, strain background (Caenorhabditis elegans) | XA7702 | Caenorhabditis Genetics Center | mdt-15(tm2182) III | |
| Strain, strain background (Escherichia coli) | OP50 | Caenorhabditis Genetics Center | OP50 | |
| Recombinant DNA reagent | k09f5.2 | Kamath et al., 2001 | RNAi | |
| Chemical compound, drug | Oleic acid C18:1 | NuChek Prep, Inc.; Elysian, Minnesota | Cat no. U-46-A | |
| Chemical compound, drug | Oil Red O (ORO) | Sigma Aldrich | Cat no. O0625 | |
| Chemical compound, drug | 5-fluoro-2’-deoxyuridine (FUDR) | Sigma Aldrich | Cat no. F0503 | |
| Chemical compound, drug | IPTG | Sigma Aldrich | Cat no. I5502 | |
| Chemical compound, drug | Carbenicillin | Sigma Aldrich | Cat no. C1389 | |
| Chemical compound, drug | Δ7 form of dafachronic acid | Frank Schroeder, Cornell University | ||
| Chemical compound, drug | DAPI stain | Thermo Scientific | Used at a concentration of 1:1000 | |
| Software, algorithm | Spot 5.2 | Nikon | Nikon Eclipse | |
| Software, algorithm | GraphPad Prism | GraphPad Software | v.9 | |
| Software, algorithm | ImageJ software | ImageJ (http://imagej.nih.gov/ij/) |
Additional files
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Supplementary file 1
Gene expression changes of selected genes in N2 PDStv compared to N2 CON adults (Ow et al., 2018 PLoS Genet 14: e1007219).
- https://cdn.elifesciences.org/articles/61459/elife-61459-supp1-v2.xlsx
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Supplementary file 2
Strains used in this study.
- https://cdn.elifesciences.org/articles/61459/elife-61459-supp2-v2.xlsx
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Supplementary file 3
Primer sequences used in this study.
- https://cdn.elifesciences.org/articles/61459/elife-61459-supp3-v2.xlsx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/61459/elife-61459-transrepform-v2.docx
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Appendix 1—figure 1—source data 1
GLP-4 is required in multiple tissues to regulate adult reproductive plasticity.
- https://cdn.elifesciences.org/articles/61459/elife-61459-app1-fig1-data1-v2.xlsx
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Appendix 1—figure 2—source data 1
Total, mitotic, and meiotic cell rows in control and PDStv animals in wild type and kri-1(ok1254).
- https://cdn.elifesciences.org/articles/61459/elife-61459-app1-fig2-data1-v2.xlsx
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Appendix 1—figure 3—source data 1
Decreased fecundity in postdauer animals results from a reduction of sperm available for self-fertilization.
- https://cdn.elifesciences.org/articles/61459/elife-61459-app1-fig3-data1-v2.xlsx
