The antimicrobial peptide defensin cooperates with tumour necrosis factor to drive tumour cell death in Drosophila
- CRUK Beatson Institute, United Kingdom
- Ecole Polytechnique Federale de Lausanne, Switzerland
- Genetic Strains Research Center, National Institute of Genetics, Japan
- University of Glasgow, United Kingdom
- CR University Grenoble Alpes, Inserm U1209, CNRS UMR5309, Immunologie Analytique des Pathologies Chroniques, France
Figures
Figure 1 with 1 supplement
def is induced in dlg mutant tumour bearing animals.
(A) Working model showing the cooperation between haemocyte-derived TNF and the immune response in the fat body in tumour cell death (Parisi et al., 2014). (B) RT-qPCR analyses showing expression of several AMPs in the fat body of dlg40.2 mutant tumour bearing larvae compared to wild-type (w1118) larvae (n = 4). (C) RT-qPCR analysis of def expression in w1118 and dlg40.2 whole larvae reared on antibiotics (n = 7). (D) RT-qPCR analysis showing def expression in larvae expressing a ctrl-IR or a dlg-IR in the posterior part of the wing disc (en>;UAS-dcr2) (n = 3). (E) Schematic representation of the def gene locus showing mutant alleles generated (UTR: Untranslated Regions, SP: Signal Peptide, PrD: Pro-Domain, Def: Mature Defensin). Statistical analysis: B-D, Student t-test, B, ***p=0.0003, C, **p=0.0074, D, *p=0.042.
Figure 1—figure supplement 1
def mediates animal survival to infection by Gram-positive bacteria.
(A) Survival of defsk3 mutant flies (n = 53), upon Gram-positive, Listeria innocua infection, is compared to wild-type (wt) (n = 55) and spatzle (spzM7) (n = 30) (the Toll ligand) mutant flies. (B) Survival of defsk3 mutant flies (n = 37), upon Gram-negative, Erwinia carotovora carotovora infection, is compared to wild-type (wt) (n = 40) and relish (relE20) (n = 10) (an Imd-pathway downstream effector) mutant flies. Statistical analysis: A, B, Log-rank test, A, ***p=0.00023, B, p=0.798.
Figure 2 with 1 supplement
Def restricts tumour growth and promotes tumour cell death.
(A-E’) Quantification of tumour volume (TV) (A) and tumour cell death (TCD) (B) in wing imaginal discs from dlg40.2 (n = 17), dlg40.2;defsk3 (n = 19) and dlg40.2;defsk4 (n = 20) mutant larvae and representative immunofluorescence images of tissues quantified (C–E’). F, RT-qPCR analysis showing def expression upon ubiquitous knockdown (dlg40.2; tub>) (n = 3). G-J’, Quantification of TV (G) and TCD (H) in wing imaginal discs from larvae ubiquitously expressing a control-IR (ctrl-IR: n = 10) or a def-IR (n = 15) and representative immunofluorescence images of tissues quantified (I–J’). K-L, Quantification of TV (K) and TCD (L) in wing imaginal discs from dlg mutant controls (dlg40.2;lpp>: n = 28), dlg;defsk3 mutants controls (dlg40.2;defsk3;lpp>: n = 31) or dlg;defsk3 mutants expressing def in the fat body (dlg40.2;defsk3,UAS-def;lpp>: n = 27). M, Effect of PBS (ctrl) or synthetic Def injection on TCD from wild-type larvae (w1118, ctrl: n = 10, Def: n = 9), dlg40.2 (ctrl: n = 10, Def: n = 10) and dlg40.2;defsk3 (ctrl: n = 11, Def: n = 18) mutant larvae. Tissues were stained with 4’,6-diamidino-2-phenylindole (DAPI, blue) for nuclei visualisation and with anti-cleaved Decapping protein 1 (Dcp1) antibody (red) to detect apoptotic cell death. Scale bars = 50 μm. Statistical analysis: A, B, K, L, One way ANOVA, *p<0.05, ****p<0.0001; F, Student t-test, ****p<0.0001, G-H, Mann-Whitney test, G, **p=0.0054, H, ****p<0.0001; M, Two way ANOVA (only relevant significant statistics are indicated), *p<0.05, ****p<0.0001.
Figure 2—figure supplement 1
Tumour suppression by Def is independent of infection and tumour cell proliferation.
(A-E’) Quantification of TV (A) and tumour proliferation (B) from dlg40.2 (n = 19), dlg40.2;defsk3 (n = 16) and dlg40.2;defsk4 (n = 17) mutant larvae and representative pictures of the corresponding tumours stained with DAPI (blue) or anti-phosphoHistone H3 (pH3) antibody to determine cell proliferation (red, C-E or white, C’–E’). F-J’, Quantification of TV (F) and TCD (G) from dlg40.2 (n = 13), dlg40.2;defsk3 (n = 13) and dlg40.2;defsk4 (n = 14) mutant larvae reared on antibiotics and representative pictures of the corresponding tumours stained with DAPI (blue) and anti-Dcp1 antibody (red) (H–J’). Scale bars = 50 μm. Statistical analysis: A, B, F, G, One way ANOVA, **p<0.01, ***p<0.001, ****p<0.0001.
Figure 3
Def from the trachea and the fat body mediates tumour cell death.
(A) RT-qPCR analysis of def expression in gut, fat body and trachea dissected from w1118 or dlg40.2 mutant larvae (n = 3). B-C, Fat body (B) and trachea (C) from dlg40.2 mutant larvae stained with DAPI (blue) and anti-Def antibody (red). D, RT-qPCR analysis of def expression in dlg40.2 mutant larvae (dlg40.2;lpp>) expressing a ctrl-IR or def-IR in the fat body (n = 4). E-H’, Quantification of TV (E) and TCD (F) in wing imaginal discs from dlg40.2 mutant larvae (dlg40.2;lpp>) expressing a ctrl-IR (n = 15) or def-IR (n = 17) in the fat body and representative immunofluorescence images of tissues quantified (G–H’). I, RT-qPCR analysis of def expression in dissected trachea from dlg40.2 mutant larvae (dlg40.2;btl>) expressing a ctrl-IR or def-IR in the trachea (n = 3). J-M’, Quantification of TV (J) and TCD (K) in wing imaginal discs from dlg40.2 mutant larvae (dlg40.2;btl>) expressing a ctrl-IR (n = 14) or def-IR (n = 16) in the trachea and representative immunofluorescence images of tissues quantified (L–M’). Tumours were stained with DAPI (blue) and anti-Dcp1 antibody (red). Scale bars = 50 μm. Statistical analysis: A, D, I, Student t-test, D, **p=0.0054, I, ****p<0.0001; E, F, J, K, Mann-Whitney test, E, F, ****p<0.0001, J, ***p=0.0009, K, ***p=0.0004.
Figure 4 with 1 supplement
Def produced by immune tissues specifically targets tumour cells.
(A-A’’) DAPI (blue), anti-Dcp1 (green) and anti-HA antibody (red) staining of dlg40.2 mutant tumour from larvae overexpressing a def-HA construct in the fat body and the trachea (dlg40.2;lpp >UAS-def-HA). B-B’’, Enlargement of inset from A’’ (white outline) showing Dcp1 (B), Def-HA (B’) and merged channels (B’’). C-C’, DAPI (blue), anti-Dcp1 (green) and anti-HA antibody (red) staining of dlg40.2 heterozygous wing disc from larvae overexpressing def-HA (dlg40.2/FM7;lpp >UAS-def-HA). D-D’’, dlg40.2 mutant tumour stained with DAPI (blue), anti-Def (red) and anti-Dcp1 (green) antibodies. E-E’’, Enlargement of inset from D’’ (white outline) showing Dcp1 (E), Def (E’) and merged channels (E’’). F, Quantification of colocalization between Def and Dcp1 staining (n = 10). G-G’, wild type (w1118) wing imaginal disc stained with DAPI (blue), anti-Def (red) and anti-Dcp1 (green) antibodies. H-H’’, High-resolution imaging of a single dying tumour cell stained with DAPI (blue), anti-Def (red) and anti-Dcp1 (green) antibodies. A, C, D, G, Scale bars = 50 μm; B, E, Scale bars = 10 μm; H, Scale bar = 2.5 μm. Statistical analysis: F, Student t-test, **p=0.0093.
Figure 4—figure supplement 1
Characterisation of UAS-def-HA expression.
(A-B) DAPI (blue) and anti-HA antibody (red) staining showing basal expression of Def-HA in trachea (A) but not in fat body (B) of UAS-def-HA larvae. C-D, DAPI (blue) and anti-HA antibody (red) staining showing expression of Def-HA in the fat body and the trachea when overexpressed with a fat body specific driver (lpp >UAS-def-HA). Scale bars = 50 μm.
Figure 5
Imd pathway activation is required for def expression and Def-mediated tumour cell death.
(A) RT-qPCR analysis of def expression from dlg40.2 and dlg40.2;imd1 mutant larvae (n = 4). B-E’, Quantification of TV (B) and TCD (C) in wing imaginal discs from dlg40.2 (n = 15) and dlg40.2;imd1 (n = 14) mutants larvae and representative immunofluorescence images of tissues quantified stained with DAPI (blue) and anti-Dcp1 antibody (red) (D–E’). F, RT-qPCR analysis showing def expression in dlg40.2 and dlg40.2; relE20 mutant animals (n = 5). G-J’, Quantification of TV (G) and TCD (H) from dlg40.2 (n = 12) and dlg40.2;relE20 (n = 10) mutant larvae and representative pictures of the corresponding tumours (I–J’). Scale bars = 50 μm. Statistical analysis: A, F, Student t-test, A, ***p<0.0003, F, ****p<0.0001; B, C, G, H, Mann-Whitney test, B, C, H, ****p<0.0001, G, **p=0.009.
Figure 6
Imd and Toll pathway activation are required in the trachea and the fat body to promote Defensin-dependent tumour cell death.
(A) RT-qPCR analysis of def expression from dlg40.2 mutant larvae (dlg40.2;btl>) expressing a ctrl-IR or imd-IR in the trachea (n = 3). B-E’, Quantification of TV (B) and TCD (C) from dlg40.2 mutant larvae (dlg40.2;btl>) expressing a ctrl-IR (n = 18) or imd-IR (n = 15) in the trachea and representative immunofluorescence images of tissues quantified stained with DAPI (blue) and anti-Dcp1 antibody (red) (D–E’). F, RT-qPCR analysis of def expression from dlg40.2 mutant larvae (dlg40.2;lpp>) expressing a ctrl-IR, an imd-IR or a myd88-IR in the fat body. G-K’, Quantification of TV (G) and TCD (H) from dlg40.2 mutant larvae (dlg40.2;lpp>) expressing a ctrl-IR, an imd-IR or a myd88-IR in the fat body and representative immunofluorescence images of tissues quantified (I–K’). L-O, Quantification of TV (L, N) and TCD (M, O) from dlg40.2 mutant larvae (dlg40.2;lpp>) expressing a ctrl-IR, an imd-IR alone or in combination with a UAS-def in the fat body (L, M) and from dlg40.2 mutant larvae (dlg40.2;lpp>) expressing a ctrl-IR, a myd88-IR alone or in combination with a UAS-def in the fat body (N, O). Scale bars = 50 μm. Statistical analysis: A, Student t-test, A, ***p=0.0001; B, C, Mann-Whitney test, B, ***p=0.0003, C, ***p=0.0002; F, G, H, L-O, One way ANOVA, *p<0.05, **p<0.01, ****p<0.0001.
Figure 7 with 3 supplements
TNF is required for PS exposure and Defensin-driven tumour cell death.
(A-A’’) DAPI (blue), Annexin V (green) and anti-Def (red) staining of dlg40.2 mutant tumours. (B-B’’) Enlargement of inset from A’’ (white outline) showing Annexin V (B), Def (B’) and merged channels (B’’). (C) Quantification of colocalisation between Def and Annexin V staining (n = 18). (D) RT-qPCR analysis showing def expression in wild-type, dlg40.2 or dlg40.2;egr3 mutants. (E- G) Annexin V (green) and DAPI (blue) staining of wing imaginal discs from larvae of the indicated genotypes. (H, I) Annexin V (green) and DAPI (blue) staining of wing imaginal discs from dlg mutant larvae (dlg40.2,hml>) expressing a ctrl-IR (n = 5) or an egr-IR (n = 9) in the haemocytes. (J) Quantification of Annexin V signal on tumours from the corresponding genotypes. (K) Quantification of TCD upon PBS (ctrl) or Def injection in wild-type (w1118, ctrl: n = 10, Def: n = 14), dlg40.2 (ctrl/Def: n = 18) or dlg40.2;egr3 (ctrl: n = 20, Def: n = 13) mutant larvae. (L) A model for Def antitumoural activity. A, E-I, Scale bars = 50 μm; B, Scale bars = 20 μm. Statistical analysis: C, Student t-test, ****p<0.0001; D, One way ANOVA, *p<0.05; J, Mann-Whitney test, ***p=0.001; K, Two way ANOVA (only relevant significant statistics are indicated), *p<0.05.
Figure 7—figure supplement 1
Def-HA does not associate to PS-negative egr-mutant tumours.
(A-B) DAPI (blue) and anti-HA antibody (red) staining showing expression of Def-HA in trachea (A) and fat body (B) of dlg40.2;egr3;lpp-gal4 >UAS def-HA larvae. C-C’, DAPI (blue), anti-Dcp1 (green) and anti-HA antibody (red) staining showing absence of Def-HA on tumours of dlg40.2;egr3;lpp-gal4 >UAS def-HA larvae. Scale bars = 50 μm.
Figure 7—figure supplement 2
TNF signalling activation does not require functional def.
(A-C’) DAPI (white), anti-Dcp1 (red) and anti-Matrix metalloproteinase 1 (Mmp1, a reporter of JNK pathway activation, green) antibody staining of wing disc from dlg40.2, dlg40.2;defsk3 and dlg40.2;defsk4 mutant larvae. D-F’, DAPI (white), Annexin V (green) and anti-Dcp1 (red) staining of tumours from dlg40.2, dlg40.2;defsk3 and dlg40.2,imd1 mutant larvae. Scale bars = 50 μm.
Figure 7—figure supplement 3
scrib mutant tumours expose PS in a TNF-dependent manner and are sensitive to Def action.
(A-A’) DAPI (blue), anti-Dcp1 (red) and Annexin V (green) staining of scrib1 mutant tumours. (B-B’) DAPI (blue), anti-Dcp1 (red) and Annexin V (green) staining of egr3;scrib1 double mutant tumours. (C-C’’) scrib1 mutant tumour stained with DAPI (blue), anti-Dcp1 (green) and anti-Def (red) antibodies. D-D’’, Enlargement of inset from C’’ (white outline) showing anti-Dcp1 (D), anti-Def (D’) and merged (D’’) channels. E, F, Quantification of tumour volume (E) and tumour cell death (F) in wing imaginal discs from scrib1 (n = 19) and defsk3;scrib1 (n = 18) mutant larvae. G-H’, representative immunofluorescence images of tissues quantified in E, F. A, B, C, G, H, Scale bars = 50 μm; D, Scale bar = 10 μm. Statistical analysis: E, F, Mann-Whitney test, E, **p=0.0015, F, ***p<0.0001.
Author response image 1
Anti-tumoural immunity in flies: the key players.
(A, B) Third instar larval imaginal wing disc overexpressing of hepwt using engrailed-gal4, UAS-rfp (eng>rfp; hepwt) (A; red). Discs where stained with AnnexinV to visualize PS exposure (B; green). (C) Assessment of apoptotic cell death in wing discs from eng>rfp; hepwtanimals injected with PBS (control) or recombinant Defensin. (D) Assessment of apoptotic cell death in wing discs from animal following overexpression of hepwt only, def only, or co-expression of both transgenes using combined eng-gal4 and lpp-gal4 drivers.
Author response image 2
Anti-tumoural immunity in flies: the key players.
(A, B) Third instar larval imaginal wing disc overexpressing of hepact using rutond-gal4 (rn>hepact).
Discs where stained with anti-Dcp1 to visualize apoptotic cell death (B; red).
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (Drosophila melanogaster) | w1118 | (Dewey et al., 2004) | BDSC: 3605; RRID:BDSC_3605 | |
| Genetic reagent (Drosophila melanogaster) | w1118 iso | (Ferreira et al., 2014) | N/A | |
| Genetic reagent (Drosophila melanogaster) | dlg40.2/FM7 | (Mendoza-Topaz et al., 2008) | Flybase_FBal0240608 | |
| Genetic reagent (Drosophila melanogaster) | FRT82B,scrib1/TM6 | (Bilder et al., 2000) | Flybase_FBal0103577 | |
| Genetic reagent (Drosophila melanogaster) | egr3 | (Igaki et al., 2002) | Flybase_FBal0147163 | |
| Genetic reagent (Drosophila melanogaster) | imd1 | (Leulier et al., 2000) | Flybase_ FBal0045906 | |
| Genetic reagent (Drosophila melanogaster) | relE20 | (Leulier et al., 2000) | DGGR: 109927; RRID:DGGR_109927 | |
| Genetic reagent (Drosophila melanogaster) | defsk3 | (Hanson et al., 2019) | N/A | |
| Genetic reagent (Drosophila melanogaster) | defsk4 | (Hanson et al., 2019) | N/A | |
| Genetic reagent (Drosophila melanogaster) | btl-gal4,UAS-RFP/CyO | Irene Miguel-Aliaga | N/A | |
| Genetic reagent (Drosophila melanogaster) | lpp-gal4/TM6B | (Brankatschk and Eaton, 2010) | N/A | |
| Genetic reagent (Drosophila melanogaster) | tub-gal4 | Bloomington Drosophila Stock Center | BDSC: 5138; RRID:BDSC_5138 | y(1) w[*]; P{w[+mC]=tubP-GAL4}LL7/TM3, Sb(4) Ser(1) |
| Genetic reagent (Drosophila melanogaster) | hmlΔ-gal4,UAS-gfp | Bruno Lemaitre | N/A | |
| Genetic reagent (Drosophila melanogaster) | en-gal4 | Bloomington Drosophila Stock Center | BDSC: 30564; RRID:BDSC_30564 | y1 w*; P{w + mW.hs=en2.4 GAL4}e16E |
| Genetic reagent (Drosophila melanogaster) | UAS-def IR | Vienna Drosophila Resource Centre | VDRC: 102437; RRID:Flybase_FBst0474306 | P{KK111656}VIE-260B |
| Genetic reagent (Drosophila melanogaster) | UAS-imd IR | Vienna Drosophila Resource Centre | VDRC: 101834; RRID:Flybase_FBst0473707 | P{KK109863}VIE-260B |
| Genetic reagent (Drosophila melanogaster) | UAS-myd88 IR | Vienna Drosophila Resource Centre | VDRC: 25402; RRID:Flybase_FBst0455868 | w1118; P{GD9716}v25402 |
| Genetic reagent (Drosophila melanogaster) | UAS-dlg IR | Vienna Drosophila Resource Centre | VDRC: 41136; RRID:Flybase_FBst0463952 | w1118; P{GD4689}v41136/TM3 |
| Genetic reagent (Drosophila melanogaster) | UAS-egr IR | Vienna Drosophila Resource Centre | VDRC: 108814; RRID:Flybase_FBst0480608 | P{KK103432}VIE-260B |
| Genetic reagent (Drosophila melanogaster) | UAS-w IR | Bloomington Drosophila Stock Center | BDSC: 25785; RRID:BDSC_25785 | y(1) v(1); P{y[+t7.7] v[+t1.8]=TRiP.JF01786}attP2 |
| Genetic reagent (Drosophila melanogaster) | UAS-def | (Tzou et al., 2000) | Flybase_FBal0145092 | |
| Genetic reagent (Drosophila melanogaster) | UAS-def-3xHA | FlyORF | FlyORF: F002467; RRID:Flybase_FBal0298643 | M{UAS-Def.ORF.3xHA.GW}ZH-86Fb |
| Genetic reagent (Drosophila melanogaster) | UAS-dcr2 | Bloomington Drosophila Stock Center | BDSC: 24650; RRID:BDSC_24650 | w[1118]; P{w[+mC]=UAS-Dcr-2.D}2 |
| Genetic reagent (Drosophila melanogaster) | spzM7 | (Neyen et al., 2014) | N/A | |
| Antibody | Anti-GFP (Chicken polyclonal) | Abcam | Cat# ab13970; RRID:AB_300798 | IF(1:4000) |
| Antibody | Anti-Def (Mouse polyclonal) | Dahua Chen (Ji et al., 2014) | N/A | IF(1:100) |
| Antibody | Anti-HA (Mouse monoclonal) | Cell Signaling Technology | Cat# 2367, RRID:AB_10691311 | IF(1:1000) |
| Antibody | Anti-dcp1 (Rabbit polyclonal) | Cell Signaling Technology | Cat# 9578, RRID:AB_2721060 | IF(1:100) |
| Antibody | Anti-phospho-Histone H3 (Ser10) (Rabbit polyclonal) | Cell Signaling Technology | Cat# 9701, RRID:AB_331535 | IF(1:100) |
| Antibody | Anti-phospho-Histone H3 (Ser28) (Rabbit polyclonal) | Cell Signaling Technology | Cat# 9713, RRID:AB_823532 | IF(1:100) |
| Antibody | Anti-Mmp1 (Mouse clonality unknown) | Developmental Studies Hybridoma Bank | Cat# 3B8D12, RRID:AB_579781 | IF(1:10) |
| Antibody | Anti-Chicken IgY Alexa 488 (Goat polyclonal) | Molecular Probes | Cat# A-11039, RRID:AB_142924 | IF(1:500) |
| Antibody | Anti-Mouse IgG Alexa 488 (Goat polyclonal) | Molecular Probes | Cat# A-11029, RRID:AB_138404 | IF(1:500) |
| Antibody | Anti-Mouse IgG Alexa 594 (Goat polyclonal) | Molecular Probes | Cat# A-11032, RRID:AB_141672 | IF(1:500) |
| Antibody | Anti-Rabbit IgG Alexa 488 (Goat polyclonal) | Molecular Probes | Cat# A-11008, RRID:AB_143165 | IF(1:500) |
| Antibody | Anti-Rabbit IgG Alexa 594 (Goat polyclonal) | Thermo Fischer Scientific | Cat# A-11037, RRID:AB_2534095 | IF(1:500) |
| Sequence-based reagent | rpl32-fwd | This paper | PCR primers | AGGCCCAAGATCGTGAAGAA |
| Sequence-based reagent | rpl32-rev | This paper | PCR primers | TGTGCACCAGGAACTTCTTGA |
| Sequence-based reagent | def-fwd | This paper | PCR primers | CTTCGTTCTCGTGGCTATCG |
| Sequence-based reagent | def-rev | This paper | PCR primers | ATCCTCATGCACCAGGACAT |
| Sequence-based reagent | def-PCR-fwd | This paper | PCR primers | TTATTGCAGAAACGGGCTCT |
| Sequence-based reagent | def-PCR-rev | This paper | PCR primers | ATGGTAAGTCGCTAACGCTAATG |
| Sequence-based reagent | def-seq | This paper | Sequencing primers | CGTGTCTTCCTGCACAGAAA |
| Sequence-based reagent | attA-fwd | This paper | PCR primers | ATGCTCGTTTGGATCTGACC |
| Sequence-based reagent | attA-rev | This paper | PCR primers | TCAAAGAGGCACCATGACCAG |
| Sequence-based reagent | cecA1-fwd | This paper | PCR primers | CTCAGACCTCACTGCAATAT |
| Sequence-based reagent | cecA1-rev | This paper | PCR primers | CCAACGCGTTCGATTTTCTT |
| Sequence-based reagent | dro-fwd | This paper | PCR primers | CGTTTTCCTGCTGCTTGCTT |
| Sequence-based reagent | dro-rev | This paper | PCR primers | GGCAGCTTGAGTCAGGTGAT |
| Sequence-based reagent | drs-fwd | This paper | PCR primers | CTCTTCGCTGTCCTGATGCT |
| Sequence-based reagent | drs-rev | This paper | PCR primers | ACAGGTCTCGTTGTCCCAGA |
| Peptide, recombinant protein | Drosophila endogenous Defensin | Bulet EIRL | N/A | |
| Peptide, recombinant protein | Drosophila Synthetic Defensin | Genepep | N/A | ATCDLLSKWNWNHTACAGHCIAKGFKGGYCNDKAVCVCRN |
| Commercial assay or kit | High Capacity cDNA Reverse Transcription Kit | Applied Biosystems | Cat# 4368813 | |
| Commercial assay or kit | PerfeCTa SYBR Green FastMix (Low ROX) | Quanta Bio | Cat# 95074–012 | |
| Commercial assay or kit | TRIzol Reagent | Thermo Fisher Scientific | Cat# 15596018 | |
| Commercial assay or kit | Turbo DNA free Kit | Life Technologies LTD | Cat# AM1907 | |
| Commercial assay or kit | High Capacity cDNA Reverse Transcription Kit | Applied Biosystems | Cat# 4368813 | |
| Chemical compound, drug | 4′,6-Diamidine-2′-phenylindole dihydrochloride (DAPI) | Sigma | Cat# D9542 | 1 μg/mL |
| Software, algorithm | Fiji | NIH | https://fiji.sc/ | |
| Software, algorithm | GraphPad Prism 6 | GraphPad | RRID:SCR_002798 | |
| Software, algorithm | 7500 Real-Time PCR Software | Applied Biosystems | RRID:SCR_014596 | |
| Software, algorithm | BatchQuantify | (Johansson et al., 2019) | https://github.com/emltwc/2018-Cell-Stem-Cell | |
| Software, algorithm | GraphPad Prism 6 | GraphPad | RRID:SCR_002798 | |
| Software, algorithm | Volocity 3D Image Analysis Software | Perkin Elmer | RRID:SCR_002668 | |
| Software, algorithm | ZEN two lite | Zeiss | RRID:SCR_013672 | |
| Other | RNasine Plus RNase Inhibitor | Promega | Cat# N261 | |
| Other | Vectashield mounting media | Vector Laboratories, Inc. | Cat# H-1000–10 | |
| Other | Annexin V, Alexa Fluor 568 conjugate | Life Technologies LTD | Cat# A13202 | 1:20 |
| Other | Annexin V, Alexa Fluor 488 conjugate | Life Technologies LTD | Cat# A13201 | 1:20 |
| Other | Penicillin-Streptomycin (10,000 U/mL) | Thermo Fisher Scientific | Cat# 15140122 | Pen: 100 IU/mL Strep: 100 mg/mL |
| Other | PfuUltra II Fusion HS DNA Polymerase | Agilent | Cat# 600670 | |
| Other | AnnexinV binding buffer | Fisher Scientific | Cat# BDB556454 |
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The antimicrobial peptide defensin cooperates with tumour necrosis factor to drive tumour cell death in Drosophila
eLife 8:e45061.
https://doi.org/10.7554/eLife.45061
