Key resources table.
| Reagent type (species) or Resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | mouse anti-Discs large | DSHB | DSHB:4F3 RRID:AB_528203 |
IHC (1:100) |
| Antibody | rabbit anti-dpERK (Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Rabbit mAb |
Cell signaling Technology | Cell signaling Technology:4370 RRID:AB_11207064 |
IHC (1:100) |
| Antibody | mouse anti-Cut | DSHB | DSHB:2B10 RRID:AB_528186 |
IHC (1:50) |
| Antibody | mouse anti- beta-galactosidase |
DSHB | DSHB:40-1a RRID:AB_528100 |
IHC (1:50) |
| Antibody | rabbit anti-Rab5 | Abcam | Abcam:ab31261 RRID:AB_882240 |
IHC (1:10000) |
| Antibody | rabbit anti-Rab7 | (Tanaka and Nakamura, 2008) | N/A | IHC (1:3000) |
| Antibody | rabbit anti-Rab11 | (Tanaka and Nakamura, 2008) | N/A | IHC (1:8000) |
| Antibody | rabbit anti-Lamp1 | Abcam | Abcam:ab30687 RRID:AB_775973 |
IHC (1:10000) |
| Antibody | mouse anti-Yan | DSHB | DSHB:8B12H9 RRID:AB_531807 |
IHC (1:200) |
| Antibody | rabbit anti-Bnl | This paper | N/A | IHC (1:1000), EIF (1:500) |
| Antibody | rabbit anti-GFP | Abcam | Abcam:ab6556 RRID:AB_305564 |
EIF (1:3000) |
| Chemical compound, drug |
Alexa Fluor 647 Phalloidin | Thermo Fisher Scientific | Cat. #: A22287 RRID:AB_2620155 |
(1:1000) |
| Genetic reagent (Drosophila melanogaster) |
{nos-Cas9}ZH-2A | Bloomington Drosophila stock center |
RRID:BDSC_54591 | |
| Genetic reagent (Drosophila melanogaster) |
UAS-CD8:GFP | Bloomington Drosophila stock center |
RRID:BDSC_5137 | |
| Genetic reagent (Drosophila melanogaster) |
UAS-nlsGFP | Bloomington Drosophila stock center |
RRID:BDSC_4776 | |
| Genetic reagent (Drosophila melanogaster) |
UAS-CD8:RFP | Bloomington Drosophila stock center |
RRID:BDSC_32218 | |
| Genetic reagent (Drosophila melanogaster) |
UAS-CD4:mIFP | Bloomington Drosophila stock center |
RRID:BDSC_64182 | |
| Genetic reagent (Drosophila melanogaster) |
lexO-nsyb:GFP1-10, UAS-CD4:GFP11 |
Bloomington Drosophila stock center |
RRID:BDSC_64315 | |
| Genetic reagent (Drosophila melanogaster) |
UAS-BtlDN | (Reichman-Fried and Shilo, 1995) | N/A | |
| Genetic reagent (Drosophila melanogaster) |
UAS-bnlRNAi | Bloomington Drosophila stock center |
RRID:BDSC_34572 | |
| Genetic reagent (Drosophila melanogaster) |
UAS-pntRNAi | Bloomington Drosophila stock center |
RRID:BDSC_35038 | |
| Genetic reagent (Drosophila melanogaster) |
UAS-cutRNAi | Bloomington Drosophila stock center |
RRID:BDSC_33967 | |
| Genetic reagent (Drosophila melanogaster) |
UAS-btlRNAi | Bloomington Drosophila stock center |
RRID:BDSC_40871 | |
| Genetic reagent (Drosophila melanogaster) |
UAS-diaRNAi | Bloomington Drosophila stock center |
RRID:BDSC_33424 | |
| Genetic reagent (Drosophila melanogaster) |
UAS-nrgRNAi | Bloomington Drosophila stock center |
RRID:BDSC_37496 | |
| Genetic reagent (Drosophila melanogaster) |
UAS-snRNAi | Bloomington Drosophila stock center |
RRID:BDSC_42615 | |
| Genetic reagent (Drosophila melanogaster) |
pnt-lacZ[07825] | Bloomington Drosophila stock center |
RRID:BDSC_11724 | |
| Genetic reagent (Drosophila melanogaster) |
sty-lacZ | Bloomington Drosophila stock center |
RRID:BDSC_11735 | |
| Genetic reagent (Drosophila melanogaster) |
bnl-LexA | (Du et al., 2017) | ||
| Genetic reagent (Drosophila melanogaster) |
bnl-Gal4 | Bloomington Drosophila stock center |
RRID:BDSC_112825 | |
| Genetic reagent (Drosophila melanogaster) |
btl-Gal4 | (Sato and Kornberg, 2002) | N/A | |
| Genetic reagent (Drosophila melanogaster) |
btl-LHG | (Roy et al., 2014) | N/A | |
| Genetic reagent (Drosophila melanogaster) |
UAS-Btl | (Roy et al., 2011a) | N/A | |
| Genetic reagent (Drosophila melanogaster) |
UAS-CD8:Cherry | (Roy et al., 2011a) | N/A | |
| Genetic reagent (Drosophila melanogaster) |
lexO-CD2:GFP | (Yagi et al., 2010) | N/A | |
| Genetic reagent (Drosophila melanogaster) |
hs-FLP; btl>y+>Gal4,
btl-mRFP1moe |
(Cabernard and Affolter, 2005) | N/A | |
| Genetic reagent (Drosophila melanogaster) |
hs-FLP; btl>y+>Gal4;
btl-mRFP1moe |
(Cabernard and Affolter, 2005) | N/A | |
| Genetic reagent (Drosophila melanogaster) |
bnl:gfpendo | This paper | N/A | Functional genomic knock-in allele by CRISPR |
| Genetic reagent (Drosophila melanogaster) |
btl:cherryendo | This paper | N/A | Functional genomic knock-in allele by CRISPR |
| Genetic reagent (Drosophila melanogaster) |
UAS-secGFP | This paper | N/A | Secreted GFP overexpression construct |
| Sequence-based reagent |
5’-GTCGGGG CCAATCGCGTCAAGCA-3’ |
This paper | N/A | Guide RNA-1 for bnl:gfpendo |
| Sequence-based reagent |
5’-AAACTGCT TGACGCGATTGGCCC-3’ |
This paper | N/A | Guide RNA-1 for bnl:gfpendo |
| Sequence-based reagent |
5’-GTCGATAT TAGCAGTAGCCTTAG-3’ |
This paper | N/A | Guide RNA-2 for bnl:gfpendo |
| Sequence-based reagent |
5’-AAACCTAA GGCTACTGCTAATAT-3’ |
This paper | N/A | Guide RNA-2 for bnl:gfpendo |
| Sequence-based reagent |
5’-GTCGCAT CACGGAGACGGTGCCGC-3’ |
This paper | N/A | Guide RNA-3 for bnl:gfpendo |
| Sequence-based reagent |
5’-AAACGCGG CACCGTCTCCGTGATG-3’ |
This paper | N/A | Guide RNA-3 for bnl:gfpendo |
| Sequence-based reagent |
5’-GTCGAGG TGTACTGATATCTAAG-3’ |
This paper | N/A | Guide RNA-1 for btl:cherryendo |
| Sequence-based reagent |
5’-AAACCTTA GATATCAGTACACCT-3’ |
This paper | N/A | Guide RNA-1 for btl:cherryendo |
| Sequence-based reagent |
5’-GTCGCGG CATCGAAAGGTCCAGAT-3’ |
This paper | N/A | Guide RNA-2 for btl:cherryendo |
| Sequence-based reagent |
5’-AAACATCT GGACCTTTCGATGCCG-3’ |
This paper | N/A | Guide RNA-2 for btl:cherryendo |
| Sequence-based reagent |
5’-CGTATGGG ATTCCGATTGTGTG-3’ |
This paper | N/A | Primer to amplify N fragment of HDR donor for bnl:gfpendo |
| Sequence-based reagent |
5’-CAGCTCCT CGCCCTTGGACATAGTG TTGCTGCTGCAATGTGGCGG-3’ |
This paper | N/A | Primer to amplify N fragment of HDR donor for bnl:gfpendo |
| Sequence-based reagent |
5’-CCGCCACA TTGCAGCAGCAACACTA TGTCCAAGGGCGAGGAGCT-3’ |
This paper | N/A | Primer to amplify mid fragment of HDR donor for bnl:gfpendo |
| Sequence-based reagent |
5’-CTGCTGA TGCTGCTGCTGCTGCCAC TCTTGTACAGCTCATC CATGCCCAG-3’ |
This paper | N/A | Primer to amplify mid fragment of HDR donor for bnl:gfpendo |
| Sequence-based reagent |
5’-CTGGGCA TGGATGAGCTGTACAAG AGTGGCAGCAGCAGCA GCATCAGCAG-3’ |
This paper | N/A | Primer to amplify C fragment of HDR donor for bnl:gfpendo |
| Sequence-based reagent |
5’-GGCTTGA GAGGTTCTTATAAAA TACTCGAG-3’ |
This paper | N/A | Primer to amplify C fragment of HDR donor for bnl:gfpendo |
| Sequence-based reagent |
5’-CTCAACTT CACCGTGACGAATGAC-3’ |
This paper | N/A | Primer to amplify N fragment of HDR donor for btl:cherryendo |
| Sequence-based reagent |
5’-GTTTCTCC ATGCGCTGACCCGT AATCAG-3’ |
This paper | N/A | Primer to amplify N fragment of HDR donor for btl:cherryendo |
| Sequence-based reagent |
5’-CTGATTAC GGGTCAGCGCATG GAGAAAC-3’ |
This paper | N/A | Primer to amplify mid fragment of HDR donor for btl:cherryendo |
| Sequence-based reagent |
5’-GGAATTCT TTTTGGTCTCCTTAT ACTACGAA-3’ |
This paper | N/A | Primer to amplify mid fragment of HDR donor for btl:cherryendo |
| Sequence-based reagent |
5’-TTCGTAGT ATAAGGAGACCAAA AAGAATTCC-3’ |
This paper | N/A | Primer to amplify C fragment of HDR donor for btl:cherryendo |
| Sequence-based reagent |
5’-GGTTCCTC TTCCATCCAAGGTTG-3’ |
This paper | N/A | Primer to amplify C fragment of HDR donor for btl:cherryendo |
| Sequence-based reagent |
5’-TTTTGGG GCCAATCGTGTGAAGC ACGGCGTGCGG-3’ |
This paper | N/A | Primer to introduce synonymous mutation at bnl-gRNA-1 recognition site in HDR donor |
| Sequence-based reagent |
5’-CCGCACG CCGTGCTTCACACGAT TGGCCCCAAAA-3’ |
This paper | N/A | Primer to introduce synonymous mutation at bnl-gRNA-1 recognition site in HDR donor |
| Sequence-based reagent |
5’-CCATATTA GCAGTAGTCTGAGCGG TAGCAGTAAC-3’ |
This paper | N/A | Primer to introduce synonymous mutation at bnl-gRNA-2 recognition site in HDR donor |
| Sequence-based reagent |
5’-GTTACTGC TACCGCTCAGACTACT GCTAATATGG-3’ |
This paper | N/A | Primer to introduce synonymous mutation at bnl-gRNA-2 recognition site in HDR donor |
| Sequence-based reagent |
5’-GGAGACG GTGCCGCAAGAGCGG GTCGAGCAG-3’ |
This paper | N/A | Primer to introduce synonymous mutation at bnl-gRNA-3 recognition site in HDR donor |
| Sequence-based reagent |
5’-CTGCTCG ACCCGCTCTTGCGGC ACCGTCTCC-3’ |
This paper | N/A | Primer to introduce synonymous mutation at bnl-gRNA-3 recognition site in HDR donor |
| Sequence-based reagent |
5’-CCGGGAA ACGTCCCCGCTGAGGT ATCAGTACACCTATAAG-3’ |
This paper | N/A | Primer to introduce synonymous mutation at btl-gRNA-1 recognition site in HDR donor |
| Sequence-based reagent |
5’-CTTATAGG TGTACTGATACCTCAG CGGGGACGTTTCCCGG-3’ |
This paper | N/A | Primer to introduce synonymous mutation at btl-gRNA-1 recognition site in HDR donor |
| sequence-based reagent |
5’-GTAGCAAT CCAAACGATGCGTAT CTGGACCTTTCGATGC-3’ |
This paper | N/A | Primer to introduce synonymous mutation at btl-gRNA-2 recognition site in HDR donor |
| Sequence-based reagent |
5’-GCATCGA AAGGTCCAGATACGCA TCGTTTGGATTGCTAC-3’ |
This paper | N/A | Primer to introduce synonymous mutation at btl-gRNA-2 recognition site in HDR donor |
| Sequence-based reagent |
5’-GTCCTGTT TAGGGGCGATAAGTGG-3’ |
This paper | N/A | Primer for bnl:gfpendo
HDR screening and sequencing |
| Sequence-based reagent |
5’-GTGTTGCG TAAGGTTAGGGCTTCG-3’ |
This paper | N/A | Primer for bnl:gfpendo
HDR screening and sequencing |
| Sequence-based reagent |
5’-GAAGCAG CACGATTTCTTCAAGAGCG-3’ |
This paper | N/A | Primer for bnl:gfpendo
HDR screening and sequencing |
| Sequence-based reagent |
5’-CGCTCTTG AAGAAATCGTGCTGCTTC-3’ |
This paper | N/A | Primer for bnl:gfpendo
HDR screening and sequencing |
| Sequence-based reagent |
5’-CGCCAGC CAGGCAAT-3’ |
This paper | N/A | Primer for bnl:gfpendo
HDR screening and sequencing |
| Sequence-based reagent |
5’-GTCCTCAA GAATGCCTCCTTGGAC-3’ |
This paper | N/A | Primer for btl:cherryendo
HDR screening and sequencing |
| Sequence-based reagent |
5’-GTCTATGA TACCTCTGACAGCTTC-3’ |
This paper | N/A | Primer for btl:cherryendo
HDR screening and sequencing |
| Sequence-based reagent |
5’-CTTCCCC GAGGGCTTCAAGTG-3’ |
This paper | N/A | Primer for btl:cherryendo
HDR screening and sequencing |
| Sequence-based reagent |
5’-CACTTGAA GCCCTCGGGGAAG-3’ |
This paper | N/A | Primer for btl:cherryendo
HDR screening and sequencing |
| Sequence-based reagent |
5’-GTTTCTCC ATGCGCTGACCCG TAATCAG-3’ |
This paper | N/A | Primer for btl:cherryendo
HDR screening and sequencing |
| Sequence-based reagent |
5’-AATTCGAG CTCGGTACAGATCTA TGCGAAGAAACCTGCGC-3’ |
This paper | N/A | Primer for UAS-SPBnl-sfGFP cloning |
| Sequence-based reagent |
5’-CCTCGCCC TTGGACATCATCGCA GATACAAGGCCCC-3’ |
This paper | N/A | Primer for UAS-SPBnl-sfGFP cloning |
| Sequence-based reagent |
5’-GGCCTTGT ATCTGCGATGATGTC CAAGGGCGAGGAG-3’ |
This paper | N/A | Primer for UAS-SPBnl-sfGFP cloning |
| Sequence-based reagent |
5’-GCCAAGC TTGCATGCCGGTACCT TACTTGTACAGCTC ATCCATGCCC-3’ |
This paper | N/A | Primer for UAS-SPBnl-sfGFP cloning |
| Recombinant DNA reagent |
pCR4Blunt-TOPO | Thermo Fisher Scientific |
N/A | |
| Recombinant DNA reagent |
pUC19 | Addgene | Addgene:50005 | |
| Recombinant DNA reagent |
pUAST | Drosophila Genomics Resource center |
DGRC:1000 | |
| Recombinant DNA reagent |
pCFD3 | (Port et al., 2014) | N/A | |
| Recombinant DNA reagent |
pCFD3-bnl_gRNA-1 | This paper | N/A | Plasmid for generation of bnl_gRNA-1 flies |
| Recombinant DNA reagent |
pCFD3-bnl_gRNA-2 | This paper | N/A | Plasmid for generation of bnl_gRNA-2 flies |
| Recombinant DNA reagent |
pCFD3-bnl_gRNA-3 | This paper | N/A | Plasmid for generation of bnl_gRNA-3 flies |
| Recombinant DNA reagent |
pCFD3-btl_gRNA-1 | This paper | N/A | Plasmid for generation of btl_gRNA-1 flies |
| Recombinant DNA reagent |
pCFD3-btl_gRNA-2 | This paper | N/A | Plasmid for generation of btl_gRNA-2 flies |
| Recombinant DNA reagent |
pCR4Blunt-TOPO -bnl:GFP-1 |
This paper | N/A | HDR donor for making bnl:gfpendo flies |
| Recombinant DNA reagent |
pCR4Blunt-TOPO -bnl:GFP-2 |
This paper | N/A | HDR donor for making bnl:gfpendo flies |
| Recombinant DNA reagent |
pCR4Blunt-TOPO -bnl:GFP-3 |
This paper | N/A | HDR donor for making bnl:gfpendo flies |
| Recombinant DNA reagent |
pCR4Blunt-TOPO- btl:Cherry-1 |
This paper | N/A | HDR donor for making btl:cherryendo flies |
| Recombinant DNA reagent |
pCR4Blunt-TOPO -btl:Cherry-2 |
This paper | N/A | HDR donor for making btl:cherryendo flies |
| Recombinant DNA reagent |
pUAST-SPBnl-sfGFP | This paper | N/A | Plasmid for generation of UAS-secGFP flies |
| Software, algorithm | Fiji (ImageJ v2.0) | (Schindelin et al., 2012) | RRID:SCR_002285 | |
| Software, algorithm | Adobe Photoshop | Adobe | RRID:SCR_014199 | |
| Software, algorithm | Microsoft Excel | Microsoft | RRID:SCR_016137 | |
| Software, algorithm | SnapGene | snapgene.com | RRID:SCR_015052 | |
| Software, algorithm | VassarStats | vassarstats.net | RRID:SCR_010263 | |
| Software, algorithm | R | r-project.org | RRID:SCR_001905 |