Bruce suppresses autophagy-regulated caspase activity and wing tissue growth in Drosophila
- Department of Genetics, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Japan
- Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Japan
- Institute for Genetic Medicine, Hokkaido University, Japan
- Institute of Life and Environmental Sciences, University of Tsukuba, Japan
- Laboratory for Cell Vigor Regulation, National Institute for Basic Biology, Japan
- Basic Biology Program, The Graduate University for Advanced Studies, SOKENDAI, Japan
Figures
Figure 1 with 2 supplements
Dcp-1 overexpression induces apoptosis independent of the apoptosome.
(A) Schematic structures and intrinsically disordered region prediction of Dcp-1 (dark cyan) and Drice (red). The predicted disordered score plot is shown. Regions wherein the score is more than 0.5 are predicted to be disordered. (B) Representative images of female wing phenotypes upon caspase overexpression using WP-Gal4. (C) Representative images of larval wing imaginal disc upon 3xHA-tagged executioner caspase (magenta) overexpression using WP-Gal4. Nuclei are visualized by Hoechst 33342 (green). Magnified images are shown in right below. Arrowheads indicate condensed nuclei. Scale bars: 50 µm (top) and 10 µm (bottom). (D) Representative images of the TUNEL staining (magenta) of larval wing imaginal discs upon 3xHA-tagged Dcp-1 overexpression using WP-Gal4. Nuclei are visualized by Hoechst 33342 (green). Magnified images are shown in right. Scale bar: 50 µm. (E) Schematic diagram of canonical apoptosis signaling in Drosophila. Panel E is reproduced from Figure 1A from Muramoto et al., 2025. (F, G) Representative images of female wing phenotypes upon Dcp-1::VENUS overexpression using WP-Gal4 with the genetic perturbation of canonical apoptosis signaling. (H) Quantification of (F) and (G). Each wing is manually classified into four (wingless, severe, mild, and intact) categories. Schematic images of the four categories are shown on the right. Sample sizes are shown in the figure.
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Figure 1—source data 1
Data used for graphs presented in Figure 1A, H.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig1-data1-v1.xlsx
Figure 1—figure supplement 1
Dcp-1 overexpression leads to its activation and the cleavage of executioner caspase substrates.
(A) Western blotting of the cleavage of C-terminally VENUS-tagged Dcp-1 overexpressed in larval wing imaginal disc using WP-Gal4. Both full-length (FL) Dcp-1::VENUS (MW: 62.7 kDa) and cleaved (CL) Dcp-1::VENUS (MW: 39.1 kDa) are detected using anti-GFP antibody. (B) Representative images of the anti-cleaved Dcp-1 antibody staining (green) in larval wing imaginal disc upon 3xHA-tagged Dcp-1 (magenta) overexpression using WP-Gal4. Nuclei are visualized by Hoechst 33342 (blue). Scale bar: 50 µm. (C) Schematic structures of fluorescent switch-on probe, GC3Ai. Caspase-mediated cleavage makes the probe fluorescent. Caspase activity is measured by the fluorescent intensity of the probe. Panel C is reproduced from Figure 2B from Shinoda and Miura, 2024. (D) Representative images of the GC3Ai fluorescent (green) in larval wing imaginal disc upon 3xHA-tagged Dcp-1 (magenta) overexpression using Nubbin-Gal4. Nuclei are visualized by Hoechst 33342 (blue). Scale bar: 50 µm. (E) Schematic structures of CD8::PARP::VENUS. Caspase-mediated cleavage generates neo-epitope of anti-cleaved PARP antibody. Caspase activity is measured by the intensity of the immunohistochemistry. Panel E is reproduced from Figure 2D from Shinoda and Miura, 2024. (F) Representative images of the anti-cleaved PARP antibody staining (magenta) in larval wing imaginal disc expressing CD8::PARP::VENUS probe (VENUS, green) and 3xHA-tagged Dcp-1 overexpression using WP-Gal4. Nuclei are visualized by Hoechst 33342 (blue). Scale bar: 50 µm.
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Figure 1—figure supplement 1—source data 1
Uncropped raw blot images of Figure 1—figure supplement 1A.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig1-figsupp1-data1-v1.zip
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Figure 1—figure supplement 1—source data 2
Uncropped annotated blot images of Figure 1—figure supplement 1A.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig1-figsupp1-data2-v1.zip
Figure 1—figure supplement 2
Dcp-1 overexpression induces apoptosis in Drosophila S2 cells.
(A) Representative images of Drosophila S2 cells transfected with mNeonGreen (mNG)-tagged wild-type (WT) or catalytically inactive (catalytic cysteine mutated to glycine; CG) Dcp-1 and Drice. Scale bars: 50 µm (left) and 5 µm (right). (B) Quantification of mNG intensity for each cell, shown as a violin plot. Quartiles are indicated by black broken lines, and the median is indicated by a red bar. (B′) Magnified view of the data in (B), with the Y-axis limited to 0–40. p-values were calculated using one-way ANOVA followed by Sidak’s multiple-comparison test for selected pairs. NS, p ≧ 0.05; †p < 0.05. Sample sizes: mNG (n = 712 cells), Dcp-1WT::mNG (n = 212 cells), Dcp-1CG::mNG (n = 380 cells), DriceWT::mNG (n = 434 cells), and DriceCG::mNG (n = 442 cells). (C) Quantification of the average mNG intensity in each field of view. Data are presented as mean ± SEM. p-values were calculated using one-way ANOVA followed by Sidak’s multiple-comparison test for selected pairs. NS, p ≧ 0.05; †p < 0.05. Sample size for each condition is 4. (D) Quantification of the mNG-positive cell number in each field of view. Data are presented as mean ± SEM. p-values were calculated using one-way ANOVA followed by Sidak’s multiple-comparison test for selected pairs. NS, p ≧ 0.05; †p < 0.05. Sample size for each condition is 4.
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Figure 1—figure supplement 2—source data 1
Data used for graphs presented in Figure 1—figure supplement 2B–D.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig1-figsupp2-data1-v1.xlsx
Figure 2 with 3 supplements
Dcp-1-enriched proximal proteins are required for Dcp-1 activation.
(A) Western blotting of expression of C-terminally V5::TurboID knocked-in tagged caspases and biotinylation patterns of their proximal proteins detected by streptavidin (SA) in larval wing imaginal discs. (B) A schematic diagram of the TurboID-mediated identification of proximal proteins. Dcp-1- and Drice-proximal proteins (Proteins C, D, E, F, and G) are promiscuously labeled in vivo with the administration of biotin. Then, biotinylated proteins are purified using NeutrAvidin magnetic beads and are subsequently analyzed by mass spectrometry. Panel B is reproduced from Figure 1F from Muramoto et al., 2025. (C) A summary of mass spectrometry analysis of proteins in the larval imaginal discs. Among 818 proteins identified, 230 proteins were detected as highly enriched proteins in Drice::V5::TurboID flies (FC [Drice::V5::TurboID/Dcp-1::V5::TurboID]>2) and 16 proteins were detected as highly enriched proteins in Dcp-1::V5::TurboID flies (FC [Dcp-1::V5::TurboID/Drice::V5::TurboID]>2). (D) Western blotting validation of the anti-CCT8 antibody in whole larval lysates without the digestive tract and fat body. The anti-CCT8 antibody detects both endogenous CCT8 (MW: 59.4 kDa) and 3xFLAG::StrepII::VENUS::StrepII (FSVS)-tagged CCT8 (MW: 91.1 kDa). (E) Western blotting of endogenous CCT8 expressed in larval wing imaginal disc. Dcp-1-proximal and Drice-proximal proteins biotinylated by C-terminally knocked-in tagged V5::TurboID are purified using NeutrAvidin. The red asterisk represents a non-specific band in the marker lane. (F) Schematic diagram of the RNAi screen targeting eight Dcp-1-enriched proximal proteins. The WP >Dcp-1::VENUS line was crossed with each UAS-RNAi line, and wing phenotypes of the F1 progeny were scored. (G) Representative images of female wing phenotypes from Dcp-1::VENUS overexpression driven by WP-Gal4 with knockdown of Dcp-1-enriched proximal proteins. ‘XX’ denotes the knocked-down gene, and ‘#1’ and ‘#2’ indicate independent RNAi lines. (H) Representative images of female wing phenotypes upon Dcp-1::VENUS overexpression driven by WP-Gal4 in the Sirt1 null mutant background. (I) Quantification of (H). Each wing is manually classified into four (wingless, severe, mild, and intact) categories. Sample sizes are shown in the figure.
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Figure 2—source data 1
Uncropped raw blot images of Figure 2A, D, E.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig2-data1-v1.zip
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Figure 2—source data 2
Uncropped annotated blot images of Figure 2A, D, E.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig2-data2-v1.zip
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Figure 2—source data 3
Data used for graphs presented in Figure 2I.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig2-data3-v1.xlsx
Figure 2—figure supplement 1
Expression patterns of Drosophila caspases in larval and adult tissues.
(A) Western blotting of expression of C-terminally V5::TurboID knocked-in tagged caspases and biotinylation patterns of their proximal proteins detected by streptavidin (SA) in larval salivary glands, fat bodies, and brains. (B) Western blotting of expression of C-terminally V5::TurboID knocked-in tagged caspases and biotinylation patterns of their proximal proteins detected by SA in adult thoraxes, ovaries, and testes.
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Figure 2—figure supplement 1—source data 1
Uncropped raw blot images of Figure 2—figure supplement 1A, B.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig2-figsupp1-data1-v1.zip
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Figure 2—figure supplement 1—source data 2
Uncropped annotated blot images of Figure 2—figure supplement 1A, B.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig2-figsupp1-data2-v1.zip
Figure 2—figure supplement 2
Comparison of Dcp-1 interactors identified by TurboID-MS and immune-affinity purification MS.
(A) Western blotting of C-terminally V5::TurboID knocked-in tagged Dcp-1 in larval wing imaginal disc. Only full-length (FL) Dcp-1::V5::TurboID (MW: 72.8 kDa), but not cleaved Dcp-1::V5::TurboID (MW: 49.2 kDa), is detected using anti-V5 antibody. (B) Pie chart of Dcp-1 interactors identified by immunoaffinity purification MS (Choutka et al., 2017). Among the 24 Dcp-1 interactors, 15 were also identified by our TurboID-MS analysis. Proteins are grouped according to their fold change relative to the control (FC [Dcp-1::V5::TurboID/w1118]).
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Figure 2—figure supplement 2—source data 1
Uncropped raw blot images of Figure 2—figure supplement 2A.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig2-figsupp2-data1-v1.zip
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Figure 2—figure supplement 2—source data 2
Uncropped annotated blot images of Figure 2—figure supplement 2A.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig2-figsupp2-data2-v1.zip
Figure 2—figure supplement 3
RNAi screening of Dcp-1-enriched proximal proteins for regulators of Dcp-1 activation.
(A) Quantification of female wing phenotypes upon knockdown of Dcp-1-enriched proximal proteins driven by WP-Gal4. ‘XX’ denotes the knocked-down gene, and ‘#1’ and ‘#2’ indicate independent RNAi lines. Each wing is manually classified into four (wingless, severe, mild, and intact) categories. Sample sizes are shown in the figure. (B) Quantification of female wing phenotypes upon Dcp-1::VENUS overexpression driven by WP-Gal4 with simultaneous knockdown of Dcp-1-enriched proximal proteins. ‘XX’ denotes the knocked-down gene, and ‘#1’ and ‘#2’ indicate independent RNAi lines. Each wing is manually classified into four (wingless, severe, mild, and intact) categories. Sample sizes are shown in the figure.
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Figure 2—figure supplement 3—source data 1
Data used for graphs presented in Figure 2—figure supplement 3A, B.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig2-figsupp3-data1-v1.xlsx
Figure 3 with 1 supplement
Debcl, Buffy, and autophagy are specifically required for Dcp-1 overexpression-induced apoptosis.
(A) Representative images of female wing phenotypes from Dcp-1::VENUS overexpression driven by WP-Gal4 with the genetic perturbation of Synr, Debcl, Buffy, and autophagy. ‘XX’ denotes the knocked-down gene, and ‘#1’ and ‘#2’ indicate independent RNAi lines. (B) Quantification of (A). ‘XX’ denotes the knocked-down gene, and ‘#1’ and ‘#2’ indicate independent RNAi lines. Each wing is manually classified into four (wingless, severe, mild, and intact) categories. Sample sizes are shown in the figure. (C) Representative images of mCherry::Atg8a puncta (autophagosome) accumulation (red) in larval wing imaginal discs upon VENUS-tagged Dcp-1 (green) overexpression using WP-Gal4. Nuclei are visualized by Hoechst 33342 (blue). Scale bars: 50 µm (top) and 10 µm (bottom). The contrast of GFP and Dcp-1::VENUS signals are adjusted differently to enhance visibility. (D) Representative images of female eye phenotypes upon Hid overexpression with the genetic perturbation of canonical apoptosis signaling. (E) Representative images of female eye phenotypes upon Hid overexpression with the genetic perturbation of Sirt1 or Fkbp59. ‘#1’ and ‘#2’ indicate independent RNAi lines. (F) Representative images of female eye phenotypes upon Hid overexpression with the genetic perturbation of Debcl, Buffy, and autophagy.
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Figure 3—source data 1
Data used for graphs presented in Figure 3B.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig3-data1-v1.xlsx
Figure 3—figure supplement 1
Knockdown of autophagy-related genes for regulators of Dcp-1 activation.
(A) Quantification of female wing phenotypes upon knockdown of Synr, Debcl, Buffy, and autophagy-related genes by WP-Gal4. ‘XX’ denotes the knocked-down gene. Each wing is manually classified into four (wingless, severe, mild, and intact) categories. Sample sizes are shown in the figure. (B) Quantification of female wing phenotypes upon knockdown of autophagy-related genes by WP-Gal4. ‘XX’ denotes the knocked-down gene. Each wing is manually classified into four (wingless, severe, mild, and intact) categories. Sample sizes are shown in the figure. (C) Quantification of female wing phenotypes upon Dcp-1::VENUS overexpression driven by WP-Gal4 with simultaneous knockdown of autophagy-related genes by. ‘XX’ denotes the knocked-down gene. Each wing is manually classified into four (wingless, severe, mild, and intact) categories. Sample sizes are shown in the figure.
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Figure 3—figure supplement 1—source data 1
Data used for graphs presented in Figure 3—figure supplement 1A–C.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig3-figsupp1-data1-v1.xlsx
Figure 4 with 2 supplements
Cleaved Dcp-1, but not Drice, interacts with Bruce.
(A) Domain structure of Bruce. LD denotes the linker domain, BIR the Baculoviral IAP repeat, WD40 a WD40 domain, β-X a β-sandwich-like fold, CBM a domain resembling the carbohydrate-binding module family 32, UBL the ubiquitin-like domain, and UBC the E2–E3 hybrid ubiquitin-conjugating domain. The region used for structural prediction using AlphaFold3 is indicated in red. (B) IBM-like sequence in Dcp-1. The propeptide cleavage site of Dcp-1 is indicated by a black dashed line; the caspase recognition motif is highlighted in black; the IBM-like sequence is highlighted in pink; and the region used for AlphaFold3 prediction is shown in red. (C) IBM-like sequence in Drice. The propeptide cleavage site of Drice is indicated by a black dashed line; the caspase recognition motif is highlighted in black; the IBM-like sequence is highlighted in pink; and the region used for AlphaFold3 prediction is shown in red. (D) Predicted complex structure of Bruce BIR and Dcp-1 IBM-like sequence. The AlphaFold3 prediction [model 0] is shown. The Bruce β-X2 and BIR domains are displayed as surfaces in green and pink, respectively. Dcp-1 is shown in stick representation and color-coded according to pLDDT scores, as indicated in the figure. See also panel (E). (E) pLDDT scores of Dcp-1 in the five predicted complex structures with Bruce. For all five AlphaFold3 models, per-residue pLDDT scores of Dcp-1 are plotted. Prediction confidence is color-coded as follows: very high (pLDDT > 90) in dark blue, confident (90 > pLDDT > 70) in light blue, low (70 > pLDDT > 50) in yellow, and very low (pLDDT < 50) in orange. (F) Predicted complex structure of Bruce BIR and Drice IBM-like sequence. The AlphaFold3 prediction [model 0] is shown. The Bruce β-X2 and BIR domains are displayed as surfaces in green and pink, respectively. Drice is shown in stick representation and color-coded according to pLDDT scores, as indicated in the figure. See also panel (G). (G) pLDDT scores of Drice in the five predicted complex structures with Bruce. For all five AlphaFold3 models, per-residue pLDDT scores of Drice are plotted. Prediction confidence is color-coded as follows: very high (pLDDT > 90) in dark blue, confident (90 > pLDDT > 70) in light blue, low (70 > pLDDT > 50) in yellow, and very low (pLDDT < 50) in orange. (H) Co-immunoprecipitation of N-terminally 3xFLAG-tagged N-terminal region of Bruce (Br, 1–900 aa; containing LD, BIR, and WD40 domains) with C-terminally myc-tagged, catalytically inactive Dcp-1 or Drice (catalytic cysteine mutated to glycine; CG) expressed in Drosophila S2 cells. N-terminally 3xFLAG-tagged mRuby3 (mRb3) was used as a control. Apoptosis was induced by cycloheximide (CHX) treatment. 3xFLAG-tagged proteins were immunoprecipitated using an anti-3xFLAG antibody. Both full-length (FL) and cleaved (CL) caspases are detected using anti-Myc antibody. For the anti-Myc blot, a contrast-enhanced (CE) image is shown below. The results displayed are representative of three biologically independent experiments. (I) Schematic diagram of wing imaginal disc samples used for co-immunoprecipitation to analyze the interaction between endogenously expressed N-terminally mStayGold::V5-tag knocked in-tagged Bruce (magenta) with C-terminally VENUS-tagged Dcp-1 (green). (J) Co-immunoprecipitation of endogenously expressed N-terminally mStayGold::V5-tag knocked in-tagged Bruce with C-terminally VENUS-tagged Dcp-1 overexpressed in wing imaginal disc using WP-Gal4. GFP overexpressed in wing imaginal disc using WP-Gal4 was used as a control. V5-tagged proteins were immunoprecipitated using an anti-V5-tag antibody. Both full-length (FL) Dcp-1::VENUS (MW: 62.7 kDa) and cleaved (CL) Dcp-1::VENUS (MW: 39.1 kDa) are detected using anti-GFP antibody. For the anti-GFP blot, a CE image is shown below. The results displayed are representative of three biologically independent experiments.
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Figure 4—source data 1
Uncropped raw blot images of Figure 4H, J.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig4-data1-v1.zip
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Figure 4—source data 2
Uncropped annotated blot images of Figure 4H, J.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig4-data2-v1.zip
Figure 4—figure supplement 1
Predicted structures of Bruce complexed with Dcp-1 and Drice IBM-like sequences.
(A) Overall view of the predicted complex structure of Bruce and Dcp-1. Model 0 is shown with the same color scheme as in Figure 4A, B. The predicted position error is shown in (A’). (B) Overall view of the predicted complex structure of Bruce and Drice. Model 0 is shown with the same color scheme as in Figure 4A, C. The predicted position error is shown in (B’). (C) Structural comparison between the experimentally determined complex of XIAP BIR3 and Smac IBM (C), and the predicted complex of Bruce BIR and Dcp-1 IBM (C’). XIAP BIR3 is shown as a pink cartoon and the Smac IBM as green sticks (PDB ID: 1G73, right panel). Bruce BIR domain is shown as a pink cartoon and the Dcp-1 IBM as dark green sticks (model 0, left panel). Zn²+ ions are shown as gray spheres. Salt bridges are indicated by black dashed lines, along with the corresponding interacting residues.
Figure 4—figure supplement 2
Expression patterns of mStayGold::V5-tag knocked-in Bruce.
(A) Schematic diagram of CRISPR/Cas9-mediated knock-in of mStayGold::C4 linker (C4)::V5 tag at the N-terminus of Bruce. A 3xP3-DsRed cassette was integrated at the endogenous TTAA site as a transformation marker. Removal of the cassette restores the genomic TTAA site, enabling scarless genome engineering. (B) Expression pattern of mStayGold (mSG)::C4::V5-tag knocked-in Bruce detected by anti-V5 tag antibody (green) in the ovariole. Nuclei are visualized using Hoechst 33342 (magenta). Scale bar: 50 µm. (C) Expression pattern of mSG::C4::V5-tag knocked-in Bruce detected by anti-V5 tag antibody (green) in the testis. F-actin is visualized using Phalloidin (magenta). Scale bar: 50 µm. (D) Western blotting of expression of mSG::C4::V5-tag knocked-in tagged Bruce in the whole adult male and female. (E) Expression pattern of mSG::C4::V5-tag knocked-in tagged Bruce detected by anti-V5 tag antibody (green) in the wing imaginal discs. Nuclei are visualized using Hoechst 33342 (magenta). Scale bar: 50 µm. (F) Expression pattern of mSG::C4::V5-tag knocked-in tagged Bruce detected by anti-V5 tag antibody (green) in the wing imaginal discs upon Bruce-RNAi using WP-Gal4. Nuclei are visualized using Hoechst 33342 (magenta). Scale bar: 50 µm.
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Figure 4—figure supplement 2—source data 1
Uncropped raw blot images of Figure 4—figure supplement 2D.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig4-figsupp2-data1-v1.zip
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Figure 4—figure supplement 2—source data 2
Uncropped annotated blot images of Figure 4—figure supplement 2D.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig4-figsupp2-data2-v1.zip
Figure 5 with 1 supplement
Bruce selectively suppresses Dcp-1 activity and wing tissue growth.
(A) Expression pattern of overexpressed myc::Bruce (green) by using WP-Gal4 in the wing imaginal discs. Nuclei are visualized using Hoechst 33342 (magenta). Scale bar: 50 µm. (B) Representative images of female wing phenotypes from Dcp-1::VENUS overexpression driven by WP-Gal4 with DIAP-1 or Bruce overexpression. (C) Quantification of (B). Each wing is manually classified into four (wingless, severe, mild, and intact) categories. Sample sizes are shown in the figure. (D) Representative images of female wing phenotypes upon Synr overexpression driven by Nubbin-Gal4 with DIAP-1 or Bruce overexpression. Arrowheads indicate wrinkled wings. (E) Representative images of female eye phenotypes upon Hid overexpression with DIAP-1 or Bruce overexpression. (F) Representative images of female wing upon Bruce or Bruce-RNAi (Bruce-i) overexpression using WP-Gal4. Scale bar: 1 mm. (G) Quantification of relative wing size normalized to the corresponding No-Gal4 control. Data are presented as mean ± SEM. p-values were calculated using one-way ANOVA followed by Dunnett’s multiple-comparison test against no UAS control. NS, p ≧ 0.05; †p < 0.05. Sample sizes are shown in the figure. (H) Representative images of female wing upon Bruce or Bruce-RNAi (Bruce-i) overexpression using Engrailed-Gal4 (En-Gal4). The gray area (labeled as A) represents the anterior part, and the green area (labeled as P) represents the posterior part of the wing. Scale bar: 1 mm. (I) Quantification of relative posterior part of wing size normalized to corresponding anterior part of wing size. Data are presented as mean ± SEM. p-values were calculated using one-way ANOVA followed by Dunnett’s multiple comparison test against no UAS control. †p < 0.05. Sample sizes are shown in the figure. (J) Representative images of female wing upon InRDN and Bruce-RNAi (Bruce-i) overexpression using WP-Gal4. Scale bar: 1 mm. (K) Quantification of relative wing size normalized to control (WP > +). Data are presented as mean ± SEM. p-values were calculated using one-way ANOVA followed by Sidak’s multiple comparison test for selected pairs. †p < 0.05. Sample sizes are shown in the figure. (L) Schematic diagram of autophagy-facilitated alternative caspase activation pathway in Drosophila mediated by autophagy–Bruce axis. Panel L is reproduced from Figure 1E from Shinoda and Miura, 2024.
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Figure 5—source data 1
Data used for graphs presented in Figure 5C, G, I, K.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig5-data1-v1.xlsx
Figure 5—figure supplement 1
Effect of Debcl or Buffy knockdown on wing tissue growth.
(A) Quantification of relative wing size normalized to the corresponding No-Gal4 control. Data are presented as mean ± SEM. p-values were calculated using one-way ANOVA followed by Dunnett’s multiple-comparison test against no UAS control. †p < 0.05. Sample sizes are shown in the figure.
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Figure 5—figure supplement 1—source data 1
Data used for graphs presented in Figure 5—figure supplement 1A.
- https://cdn.elifesciences.org/articles/109876/elife-109876-fig5-figsupp1-data1-v1.xlsx
Tables
Appendix 1—key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Drosophila melanogaster) | Dcp-1 | FlyBase | FLYB: FBgn0010501 | |
| Gene (D. melanogaster) | Dcp-1-PA | FlyBase | FLYB: FBpp0071971 | |
| Gene (D. melanogaster) | Drice | FlyBase | FLYB: FBgn0019972 | |
| Gene (D. melanogaster) | Drice-PA | FlyBase | FLYB: FBpp0084848 | |
| Gene (D. melanogaster) | CCT8 | FlyBase | FLYB: FBgn0284436 | |
| Gene (D. melanogaster) | CCT8-PA | FlyBase | FLYB: FBpp0087764 | |
| Gene (D. melanogaster) | Bruce | FlyBase | FLYB: FBgn0266717 | |
| Gene (D. melanogaster) | Bruce-PB | FlyBase | FLYB: FBpp0304070 | |
| Strain, strain background (Escherichia coli) | BL21 (B strain) | TaKaRa | 9126 | |
| Genetic reagent (D. melanogaster) | w1118 | Shinoda et al., 2023 | FLYB: FBal0018186 | Maintained in Masayuki Miura lab |
| Genetic reagent (D. melanogaster) | WP-Gal4 | Bloomington Drosophila Stock Center | RRID:BDSC_49828 | |
| Genetic reagent (D. melanogaster) | UAS-Dcp-1::3xHA | FlyORF | FlyORF ID: F003889; FLYB: FBal0350652 | |
| Genetic reagent (D. melanogaster) | UAS-Drice::3xHA | FlyORF | FlyORF ID: F002751; FLYB: FBal0303612 | |
| Genetic reagent (D. melanogaster) | UAS-Dcp-1::VENUS | https://doi.org/10.1016/j.neuron.2011.09.031 | FLYB: FBal0269199 | Gift from Dr. Graeme W. Davis |
| Genetic reagent (D. melanogaster) | Df(3L)H99 | Kyoto Stock Center | DGRC Number: 106395; FLYB: FBst0305064 | |
| Genetic reagent (D. melanogaster) | UAS-LacZ-RNAi | https://doi.org/10.1038/78531 | FLYB: FBal0196940 | Gift from Dr. Richard Carthew |
| Genetic reagent (D. melanogaster) | UAS-p35 | Bloomington Drosophila Stock Center | RRID:BDSC_5072 | Gift from Dr. Bruce Hay |
| Genetic reagent (D. melanogaster) | UAS-miRHG | https://doi.org/10.1016/j.cub.2010.01.060 | FLYB: FBti0301040 | Gift from Dr. Chung-Hong Chen |
| Genetic reagent (D. melanogaster) | UAS-Drice-RNAi | Vienna Drosophila Resource Center | VDRC ID: 28065; FLYB: FBst0457273 | |
| Genetic reagent (D. melanogaster) | UAS-Dark-RNAi | Vienna Drosophila Resource Center | VDRC ID: 100405; FLYB: FBst0472278 | |
| Genetic reagent (D. melanogaster) | UAS-Dronc-RNAi | Vienna Drosophila Resource Center | VDRC ID: 23035; FLYB: FBst0454804 | |
| Genetic reagent (D. melanogaster) | UAS-DroncDN::EGFP | https://doi.org/10.1074/jbc.M002935200 | FLYB: FBal0122050 | Gift from Dr. Helena Richardson |
| Genetic reagent (D. melanogaster) | Nubbin-Gal4 | Bloomington Drosophila Stock Center | RRID:BDSC_25754 | Gift from Dr. Sa Kan Yoo |
| Genetic reagent (D. melanogaster) | UAS-GC3Ai | Bloomington Drosophila Stock Center | RRID:BDSC_84343 | Gift from Dr. Sa Kan Yoo |
| Genetic reagent (D. melanogaster) | UAS-CD8::PARP::VENUS | Kyoto Stock Center | DGRC Number: 116295; FLYB: FBti0182700 | Gift from Dr. Darren W. Williams |
| Genetic reagent (D. melanogaster) | Dcp-1::V5::TurboID | Shinoda et al., 2019 | FLYB: FBal0356245 | |
| Genetic reagent (D. melanogaster) | Drice::V5::TurboID | Shinoda et al., 2019 | FLYB: FBal0356244 | |
| Genetic reagent (D. melanogaster) | Dronc::V5::TurboID | Shinoda et al., 2019 | FLYB: FBal0356243 | |
| Genetic reagent (D. melanogaster) | CCT8::3xFLAG::StrepII::VENUS::StrepIICPTI003689 | Kyoto Stock Center | DGRC Number: 115440; FLYB: FBst0325427 | |
| Genetic reagent (D. melanogaster) | UAS-white-RNAi | NIG-FLY | RRID:BDSC_33644 | |
| Genetic reagent (D. melanogaster) | UAS-p23-RNAi #1 | Vienna Drosophila Resource Center | VDRC ID: 15309; FLYB: FBst0451796 | |
| Genetic reagent (D. melanogaster) | UAS-p23-RNAi #2 | NIG-FLY | RRID:BDSC_62911 | |
| Genetic reagent (D. melanogaster) | UAS-CCT8-RNAi #1 | Vienna Drosophila Resource Center | VDRC ID: 103905; FLYB: FBst0475763 | |
| Genetic reagent (D. melanogaster) | UAS-CCT8-RNAi #2 | NIG-FLY | NIG-FLY ID: 8258R-2; FLYB: FBst1091918 | |
| Genetic reagent (D. melanogaster) | UAS-NudC-RNAi #1 | NIG-FLY | RRID:BDSC_52980 | |
| Genetic reagent (D. melanogaster) | UAS-NudC-RNAi #2 | Bloomington Drosophila Stock Center | RRID:BDSC_33382 | |
| Genetic reagent (D. melanogaster) | UAS-Fkbp59-RNAi #1 | Vienna Drosophila Resource Center | VDRC ID: 21862; FLYB: FBst0454273 | |
| Genetic reagent (D. melanogaster) | UAS-Fkbp59-RNAi #2 | Bloomington Drosophila Stock Center | RRID:BDSC_28349 | |
| Genetic reagent (D. melanogaster) | UAS-CCT2-RNAi #1 | Bloomington Drosophila Stock Center | RRID:BDSC_34711 | |
| Genetic reagent (D. melanogaster) | UAS-CCT2-RNAi #2 | Vienna Drosophila Resource Center | VDRC ID: 108615; FLYB: FBst0480425 | |
| Genetic reagent (D. melanogaster) | UAS-Sgt1-RNAi #1 | Vienna Drosophila Resource Center | VDRC ID: 330722; FLYB: FBst0492341 | |
| Genetic reagent (D. melanogaster) | UAS-Sgt1-RNAi #2 | Bloomington Drosophila Stock Center | RRID:BDSC_34605 | |
| Genetic reagent (D. melanogaster) | UAS-Sirt1-RNAi #1 | Vienna Drosophila Resource Center | VDRC ID: 105502; FLYB: FBst0477328 | |
| Genetic reagent (D. melanogaster) | UAS-Sirt1-RNAi #2 | Vienna Drosophila Resource Center | VDRC ID: 23201; FLYB: FBst0454875 | |
| Genetic reagent (D. melanogaster) | UAS-CG3226-RNAi #1 | Vienna Drosophila Resource Center | VDRC ID: 47035; FLYB: FBst0467028 | |
| Genetic reagent (D. melanogaster) | UAS-CG3226-RNAi #2 | Bloomington Drosophila Stock Center | RRID:BDSC_32875 | |
| Genetic reagent (D. melanogaster) | Sirt12A-7-11 | Bloomington Drosophila Stock Center | RRID:BDSC_8838 | |
| Genetic reagent (D. melanogaster) | Sirt14.5 | Bloomington Drosophila Stock Center | RRID:BDSC_32568 | |
| Genetic reagent (D. melanogaster) | UAS-Synr-RNAi #1 | Ikegawa et al., 2023 | FLYB: FBal0390889 | Gift from Dr. Sa Kan Yoo |
| Genetic reagent (D. melanogaster) | UAS-Synr-RNAi #2 | Ikegawa et al., 2023 | FLYB: FBal0390890 | Gift from Dr. Sa Kan Yoo |
| Genetic reagent (D. melanogaster) | UAS-Debcl-RNAi | Bloomington Drosophila Stock Center | RRID:BDSC_27083 | Gift from Dr. Sa Kan Yoo |
| Genetic reagent (D. melanogaster) | UAS-Buffy-RNAi | Bloomington Drosophila Stock Center | RRID:BDSC_29608 | Gift from Dr. Sa Kan Yoo |
| Genetic reagent (D. melanogaster) | UAS-Atg2-RNAi | Bloomington Drosophila Stock Center | RRID:BDSC_27706 | Gift from Dr. Sa Kan Yoo |
| Genetic reagent (D. melanogaster) | UAS-Atg8a-RNAi | Bloomington Drosophila Stock Center | RRID:BDSC_28989 | Gift from Dr. Sa Kan Yoo |
| Genetic reagent (D. melanogaster) | Act-mCherry::Atg8a | Dr. Eric Baehrecke; Ikegawa et al., 2023 | NA | Gift from Dr. Sa Kan Yoo |
| Genetic reagent (D. melanogaster) | UAS-GFP | Bloomington Drosophila Stock Center | RRID:BDSC_1521 | |
| Genetic reagent (D. melanogaster) | GMR-Hid | https://doi.org/10.1101/gad.9.14.1694 | FLYB: FBal0265023 | Gift from Dr. Hermann Steller |
| Genetic reagent (D. melanogaster) | GBS-Gal4 | https://doi.org/10.1242/dev.00624 | FLYB: FBal0152222 | Gift from Dr. Yasushi Hiromi |
| Genetic reagent (D. melanogaster) | UAS-Dcp-1-RNAi | Vienna Drosophila Resource Center | VDRC ID: 107560; FLYB: FBst0479380 | |
| Genetic reagent (D. melanogaster) | UAS-FIP200-RNAi | Bloomington Drosophila Stock Center | RRID:BDSC_36918 | Gift from Dr. Naonobu Fujita |
| Genetic reagent (D. melanogaster) | UAS-Atg9-RNAi | Bloomington Drosophila Stock Center | RRID:BDSC_34901; FLYB: FBal0263259 | Gift from Dr. Naonobu Fujita |
| Genetic reagent (D. melanogaster) | UAS-Atg5-RNAi | NIG-FLY | NIG-FLY ID: 1643R-2; FLYB: FBst1087795 | Gift from Dr. Naonobu Fujita |
| Genetic reagent (D. melanogaster) | UAS-Stx17-RNAi | Vienna Drosophila Resource Center | VDRC ID: 108825; FLYB: FBst0480619 | Gift from Dr. Naonobu Fujita |
| Genetic reagent (D. melanogaster) | mStayGold::C4 linker::V5::Bruce3xP3-DsRed | This paper; Deposited at Kyoto Stock Center | DGRC Number: 119866 | Novel transgenic fly line generated in this study; see ‘Materials and methods’ section; available from the Kyoto Stock Center (DGRC Stock Number 119866) |
| Genetic reagent (D. melanogaster) | UAS-Bruce-RNAi | Vienna Drosophila Resource Center | VDRC ID: 107620; FLYB: FBst0479439 | |
| Genetic reagent (D. melanogaster) | UAS-myc::Bruce | Domingues and Ryoo, 2012 | FLYB: FBal0268097 | Gift from Dr. Hyung Don Ryoo |
| Genetic reagent (D. melanogaster) | UAS-GFP | Bloomington Drosophila Stock Center | RRID:BDSC_1522 | |
| Genetic reagent (D. melanogaster) | UAS-DIAP-1::VENUS | Kyoto Stock Center | DGRC Number: 118637; FLYB: FBal0244058 | |
| Genetic reagent (D. melanogaster) | UAS-Synr::3xHA | FlyORF | FlyORF ID: F003800; FLYB: FBal0350567 | Gift from Dr. Sa Kan Yoo |
| Genetic reagent (D. melanogaster) | Engrailed-Gal4 | Bloomington Drosophila Stock Center | RRID:BDSC_30564 | |
| Genetic reagent (D. melanogaster) | UAS-InRDN | Bloomington Drosophila Stock Center | RRID:BDSC_8253 | |
| Cell line (D. melanogaster) | S2 | RIKEN BRC | Cat#: RCB1153; RRID:CVCL_Z232 | Maintained in Masayuki Miura lab |
| Antibody | anti-GFP (rabbit polyclonal) | MBL | Cat#: 598; RRID:AB_591816 | WB (1:2000) |
| Antibody | anti-V5 tag (mouse monoclonal) | Invitrogen | Cat#: R960-25; RRID:AB_2556564 | WB (1:1000), IHC (1:100) |
| Antibody | anti-FLAG M2 (mouse monoclonal) | Sigma | Cat#: F1804; RRID:AB_262044 | WB (1:5000) |
| Antibody | anti-CCT8 (rabbit polyclonal) | This paper | NA | WB (1:5000); novel antibody generated in this study; see ‘Materials and methods’ section; available from the corresponding authors upon reasonable request |
| Antibody | anti-Myc-Tag (71D10) (rabbit monoclonal) | CST | Cat#: 2278; RRID:AB_490778 | WB (1:5000) |
| Antibody | anti-3xDYKDDDDK Tag (3xFLAG Tag) (E7C5T) (rabbit monoclonal) | CST | Cat#: 87537; RRID:AB_3720194 | WB (1:5000) |
| Antibody | anti-alpha tubulin (DM1A) (mouse monoclonal) | Sigma | Cat#: T9026; RRID:AB_477593 | WB (1:10,000) |
| Antibody | anti-Histone H3 (1B1B2) (mouse monoclonal) | CST | Cat#: 14269; RRID:AB_2756816 | WB (1:10,000) |
| Antibody | anti-Actin (AC-40) (mouse monoclonal) | Sigma | Cat#: A4700; RRID:AB_476730 | WB (1:5000) |
| Antibody | anti-alpha tubulin (rabbit polyclonal) | Abcam | Cat#: ab15246; RRID:AB_301787 | WB (1:2000) |
| Antibody | anti-eEF2 (rabbit polyclonal) | Abcam | Cat#: ab33523; RRID:AB_732081 | WB (1:2000) |
| Antibody | anti-HA (Y11) (rabbit polyclonal) | Santa Cruz | Cat#: sc-805; RRID:AB_631618 | IHC (1:100) |
| Antibody | anti-HA (12CA5) (mouse monoclonal) | Abcam | Cat#: ab1424; RRID:AB_301017 | IHC (1:200) |
| Antibody | anti-cleaved Drosophila Dcp-1 (Asp215) (rabbit polyclonal) | CST | Cat#: 9578S; RRID:AB_2721060 | IHC (1:200) |
| Antibody | anti-cleaved PARP (Asp214) (D64E10) (rabbit monoclonal) | CST | Cat#: 5625S; RRID:AB_10699459 | IHC (1:200) |
| Antibody | anti-Myc-tag (mouse monoclonal) | Invitrogen | Cat#: R950-25; RRID:AB_2556560 | IHC (1:500) |
| Antibody | anti-rabbit IgG HRP-conjugated (goat polyclonal) | CST | Cat#: 7074S; RRID:AB_2099233 | WB (1:5000) |
| Antibody | anti-mouse IgG HRP-conjugated (goat polyclonal) | Promega | Cat#: W402B; RRID:AB_430834 | WB (1:5000) |
| Antibody | anti-rabbit IgG Cy3-conjugated (donkey polyclonal) | Jackson Immuno Research | Cat#: 711-165-152; RRID:AB_2307443 | IHC (1:250) |
| Antibody | anti-mouse IgG Alexa Fluor 647-conjugated (donkey polyclonal) | Thermo Fisher Scientific | Cat#: A-31571; RRID:AB_162542 | IHC (1:100) |
| Antibody | anti-rabbit IgG Alexa Fluor 488-conjugated (donkey polyclonal) | Thermo Fisher Scientific | Cat#: A-21206; RRID:AB_2535792 | IHC (1:200) |
| Antibody | anti-rabbit IgG Alexa Fluor 647-conjugated (donkey polyclonal) | Thermo Fisher Scientific | Cat#: A-31573; AB_2536183 | IHC (1:200) |
| Antibody | anti-mouse IgG Alexa Fluor 488-conjugated (donkey polyclonal) | Thermo Fisher Scientific | Cat#: A-21202; RRID:AB_141607 | IHC (1:500 (Myc), 1:100 (V5)) |
| Recombinant DNA reagent | pAc5-STABLE2-neo (plasmid) | Addgene | Cat#: 32426; RRID:Addgene_32426 | |
| Recombinant DNA reagent | pAc5-myc-mNeonGreen (plasmid) | This paper | Novel plasmid generated in this study; see ‘Materials and methods’ section; available from the corresponding authors upon reasonable request | |
| Recombinant DNA reagent | pAc5-Dcp-1WT-myc-mNeonGreen (plasmid) | This paper | Novel plasmid generated in this study; see ‘Materials and methods’ section; available from the corresponding authors upon reasonable request | |
| Recombinant DNA reagent | pAc5-Dcp-1CG-myc-mNeonGreen (plasmid) | This paper | Novel plasmid generated in this study; see ‘Materials and methods’ section; available from the corresponding authors upon reasonable request | |
| Recombinant DNA reagent | pAc5-DriceWT-myc-mNeonGreen (plasmid) | This paper | Novel plasmid generated in this study; see ‘Materials and methods’ section; available from the corresponding authors upon reasonable request | |
| Recombinant DNA reagent | pAc5-DriceCG-myc-mNeonGreen (plasmid) | This paper | Novel plasmid generated in this study; see ‘Materials and methods’ section; available from the corresponding authors upon reasonable request | |
| Recombinant DNA reagent | pCold I DNA (plasmid) | TaKaRa | Cat#: 3361 | |
| Recombinant DNA reagent | pCold I-EcoRI-CCT8-EcoRI (plasmid) | This paper | Novel plasmid generated in this study; see ‘Materials and methods’ section; available from the corresponding authors upon reasonable request | |
| Recombinant DNA reagent | pAc5-Dcp-1CG-myc (plasmid) | This paper | Novel plasmid generated in this study; see ‘Materials and methods’ section; available from the corresponding authors upon reasonable request | |
| Recombinant DNA reagent | pAc5-DriceCG-myc (plasmid) | This paper | Novel plasmid generated in this study; see ‘Materials and methods’ section; available from the corresponding authors upon reasonable request | |
| Recombinant DNA reagent | pAc5-3xFLAG-mRuby3 (plasmid) | This paper | Novel plasmid generated in this study; see ‘Materials and methods’ section; available from the corresponding authors upon reasonable request | |
| Recombinant DNA reagent | pAc5-Bruce1-900aa-3xFLAG (plasmid) | This paper | Novel plasmid generated in this study; see ‘Materials and methods’ section; available from the corresponding authors upon reasonable request | |
| Recombinant DNA reagent | pDCC6 (plasmid) | Addgene; Gokcezade et al., 2014 | Cat#: 59985; RRID:Addgene_59985 | |
| Recombinant DNA reagent | pDCC6-Bruce-sgRNA (plasmid) | This paper | Novel plasmid generated in this study; see ‘Materials and methods’ section; available from the corresponding authors upon reasonable request | |
| Recombinant DNA reagent | pBac[3xP3-DsRed_polyA_Scarless_TK] (plasmid) | Shinoda et al., 2019 | ||
| Recombinant DNA reagent | pRSETB/mStayGold(c4) (plasmid) | Addgene; Ando et al., 2024 | Cat#: 212018; RRID:Addgene_212018 | |
| Recombinant DNA reagent | pBac-mStayGold-c4-V5-Bruce-donor-3xP3-dsRed-polyA (plasmid) | This paper | Novel plasmid generated in this study; see ‘Materials and methods’ section; available from the corresponding authors upon reasonable request | |
| Commercial assay or kit | MycoStrip | InvivoGen | Cat#: rep-mys-10 | |
| Commercial assay or kit | Effectene Transfection Reagent | QIAGEN | Cat#: 301427 | |
| Commercial assay or kit | FG-NeutrAvidin beads | Tamagawa Seiki | Cat#: TAS8848 N1171 | |
| Commercial assay or kit | QIAexpress Ni-NTA Fast Start Kit | QIAGEN | Cat#: 30600 | |
| Commercial assay or kit | Anti-DDDDK-tag mAb-Magnetic Agarose | MBL | Cat#: M185-10R | |
| Commercial assay or kit | Anti-V5-tag mAb-Magnetic Agarose | MBL | Cat#: M167-10 | |
| Commercial assay or kit | ApopTag Red in situ apoptosis detection kit | Millipore | Cat#: S7165 | |
| Chemical compound, drug | (+)-Biotin | Wako | Cat#: 029-08713 | |
| Chemical compound, drug | Streptavidin HRP-conjugated | Invitrogen | Cat#: SA10001 | WB (1:10,000) |
| Chemical compound, drug | Cycloheximide | Sigma | Cat#: C7698 | |
| Chemical compound, drug | SlowFade Gold antifade reagent | Invitrogen | Cat#: S36939 | |
| Chemical compound, drug | Hoechst 33342 | Invitrogen | Cat#: H3570 | IHC (8 µM) |
| Chemical compound, drug | Phalloidin-iFluor633 | Abcam | Cat#: ab176758 | IHC (1:1000) |
| Chemical compound, drug | Euparal | Waldeck | Cat#: 3C-239 | |
| Software, algorithm | FlyBase | FlyBase | RRID:SCR_006549 | https://flybase.org/ |
| Software, algorithm | PSIPRED | PSIPRED | RRID:SCR_010246 | http://bioinf.cs.ucl.ac.uk/psipred/ |
| Software, algorithm | flyCRISPR | Gratz et al., 2014 | http://targetfinder.flycrispr.neuro.brown.edu/ | |
| Software, algorithm | Fiji (ImageJ) | NIH Image | RRID:SCR_002285 | https://imagej.net/software/fiji/ |
| Software, algorithm | Proteome Discoverer (PD) 2.2 | Thermo Fisher Scientific | RRID:SCR_014477 | |
| Software, algorithm | AlphaFold3 | Abramson et al., 2024 | https://alphafoldserver.com/ | |
| Software, algorithm | PyMOL | Schrödinger, LLC | RRID:SCR_000305 | |
| Software, algorithm | Drosophila wing digital twin | Datamarkin | https://datamarkin.com/models/drosophila-wings-beta | |
| Software, algorithm | GraphPad Prism 8 | GraphPad | RRID:SCR_002798 |
Additional files
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Supplementary file 1
Dcp-1 and Drice proximal protein lists.
Lists of the identified Dcp-1 proximal proteins (abundance ratio (Dcp-1::V5::TurboID/Drice::V5::TurboID) >2), Drice proximal proteins (abundance ratio (Drice::V5::TurboID/Dcp-1::V5::TurboID) >2), and candidates for RNAi screening (abundance ratio (Dcp-1::V5::TurboID/Drice::V5::TurboID) >1.5, abundance ratio (Dcp-1::V5::TurboID/Control) >10, PSM ≧2). The UniProt accessions, gene descriptions, abundance ratios, and # PSMs are listed.
- https://cdn.elifesciences.org/articles/109876/elife-109876-supp1-v1.xlsx
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Supplementary file 2
Comparison of Dcp-1 interactors.
List of previously reported Dcp-1 interactors and comparison with Dcp-1 proximal proteins identified by TurboID-MS. The UniProt accessions, gene descriptions, abundance ratios, and # PSMs are listed.
- https://cdn.elifesciences.org/articles/109876/elife-109876-supp2-v1.xlsx
-
Supplementary file 3
Fly stock list.
List of fly strains. Genotypes, maps, source or references, and stock numbers are listed.
- https://cdn.elifesciences.org/articles/109876/elife-109876-supp3-v1.xlsx
-
Supplementary file 4
Detailed genotypes.
Detailed genotypes and their related figure # are presented.
- https://cdn.elifesciences.org/articles/109876/elife-109876-supp4-v1.xlsx
-
Supplementary file 5
Liquid chromatography (LC) settings.
Gradient settings for LC analysis are presented.
- https://cdn.elifesciences.org/articles/109876/elife-109876-supp5-v1.xlsx
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Supplementary file 6
Tandem mass spectrometry (MS/MS) settings.
Settings used for the MS/MS analysis are presented.
- https://cdn.elifesciences.org/articles/109876/elife-109876-supp6-v1.xlsx
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Supplementary file 7
Proteome Discoverer 2.2 settings.
Proteome Discoverer 2.2 settings for proteomics analysis are presented.
- https://cdn.elifesciences.org/articles/109876/elife-109876-supp7-v1.xlsx
-
MDAR checklist
- https://cdn.elifesciences.org/articles/109876/elife-109876-mdarchecklist1-v1.docx
Download links
A two-part list of links to download the article, or parts of the article, in various formats.
Downloads (link to download the article as PDF)
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Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
Bruce suppresses autophagy-regulated caspase activity and wing tissue growth in Drosophila
eLife 15:RP109876.
https://doi.org/10.7554/eLife.109876.3
