Key resources
| Reagent or resource . | Source . | Identifier . |
|---|---|---|
| Antibodies | ||
| Alexa Fluor 568 phalloidin (1:500 dilution) | Thermo Fisher Scientific | Cat#A12380 |
| Mouse monoclonal anti-Arm (1:75 dilution) | DSHB | DSHB Cat#N2.7A1; RRID: AB_528089 |
| Rat monoclonal anti-Ecad (1:50 dilution) | DSHB | DSHB Cat#DCAD2; RRID: AB_528120 |
| Rabbit polyclonal anti-GFP (1:300 dilution) | Invitrogen | Cat#A10260; RRID: AB_2534022 |
| Rabbit polyclonal anti-mCherry (1:500 dilution) | Novus Bio- logicals | Cat#NBP2-25157 |
| Mouse monoclonal anti-Cactus (1:10 dilution) | DSHB | DSHB Cat#anti-cactus 3H12; RRID: AB_528109 |
| Mouse monoclonal anti-Dorsal (1:10 dilution) | DSHB | DSHB Cat#anti-dorsal 7A4; RRID: AB_528204 |
| Mouse monoclonal anti-FasIII (1:10 dilution) | DSHB | DSHB Cat#7G10 anti-Fasciclin III; RRID: AB_528238 |
| Mouse monoclonal anti-Dlg (1:10 dilution) | DSHB | DSHB Cat#4F3 anti-discs large; RRID: AB_528238 |
| Mouse monoclonal anti-Crb (1:10 dilution) | DSHB | DSHB Cat#cq4; RRID: AB_528181 |
| Rabbit anti-aPKC (1:200 dilution) | Santa Cruz Biotech-nology | Cat#sc-216 |
| Chemicals, peptides, and recombinant proteins | ||
| Hoechst 33342 | Sigma-Aldrich | Cat# 14533 |
| Experimental models: organisms/strains | ||
| D. melanogaster: c306-Gal4: P{w[+mW.hs]=GawB}c306, w[1118] | BDSC | FBti0003935 RRID: BDSC_3743 |
| D. melanogaster: y[1] w[*]; P{y[+t*] w[+mC]=UAS-Lifeact-GFP}VIE-260B | Hatan et al. (2011) | FBtp0064437 |
| D. melanogaster: w[1118] | BDSC | FBst0003605 RRID: BDSC_3605 |
| D. melanogaster: RNAi of Cactin: w1118; P{GD9200}v32718 | Vienna Drosophila Resource Center | FBti0098576 RRID: v32718 |
| D. melanogaster: RNAi of Cactin w1118; P{GD9200}v32719 | Vienna Drosophila Resource Center | FBti0098576 RRID: v32719 |
| D. melanogaster: RNAi of Cactin P{KK100507}VIE-260B | Vienna Drosophila Resource Center | FBti0116514 RRID: v106979 |
| D. melanogaster: P{tubP-GAL80ts}2 | BDSC | FBti0027797 RRID: BDSC_7017 |
| D. melanogaster: UAS-Cactin | This work | N/A |
| D. melanogaster: UAS-Cactin-RFP | This work | N/A |
| D. melanogaster: P{UAS-mCherry.CAAX.S}2 | BDSC | FBti0164914 RRID: BDSC_59021 |
| D. melanogaster: Fruitless-Gal4 | Borensztejn et al. (2013) | N/A |
| D. melanogaster: Upd-Gal4: P{GawB}E132, w* | BDSC | FBti0002638 RRID: BDSC_26796 |
| D. melanogaster: RNAi of white: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.HMS00017}attP2 | BDSC | FBst0033623 RRID: BDSC_33623 |
| D. melanogaster: Hsflp; AY-Gal4, UAS-GFP | D. Montell lab stock | N/A |
| D. melanogaster: w[1118]; P{10XStat92E-GFP}1 | BDSC | FBst0026197 RRID: BDSC_26197 |
| D. melanogaster: UAS-cactus: M{UAS-cact.ORF.3xHA}ZH-86Fb | FlyORF | FBti0153388 |
| D. melanogaster: w[*]; P{w[+mC]=sqh-mCherry.M}3 | BDSC | FBst0059024 RRID: BDSC_59024 |
| D. melanogaster: y1 w*; P{UASp-YFP.Rab5}02 | BDSC | FBti0100788 RRID: BDSC_24616 |
| D. melanogaster: y1 w*; P{UASp-YFP.Rab7}vig218 | BDSC | FBst0023270 RRID: BDSC_23270 |
| D. melanogaster: y1 w*; P{UASp-YFP.Rab11}sra32/TM3, Sb1 | BDSC | FBst0009790 RRID: BDSC_9790 |
| D. melanogaster: apkc mutant: y1 w67c23; P{lacW}aPKCk06403/CyO | BDSC | FBst0010622 RRID: BDSC_10622 |
| D. melanogaster: EGFP-aPKC | Chen et al. (2018) | FBti0211865 |
| D. melanogaster: UAS-GrabFP-Apical: M{lexAop-UAS-GrabFP.A.Int.mCh}ZH-86Fb | BDSC | FBti0187198 RRID: BDSC_68178 |
| D. melanogaster: UAS-GrabFP-Basal: M{lexAop-UAS-GrabFP.B.Int.mCh}ZH-86Fb | BDSC | FBst0068175 RRID: BDSC_68175 |
| D. melanogaster: UAS-Rac-FRET: P{w[+mC]=UAS-YPet-PAK-RAC-CFP.FRET}3 | BDSC | FBst0031431 RRID: BDSC_31431 |
| D. melanogaster: Rac2 mutant: Rac2[Delta] ry[506] | BDSC | FBst0006675 RRID: BDSC_6675 |
| D. melanogaster: Mtl mutant: y[1] w[*]; P{ry[+t7.2]=neoFRT}82B Mtl[Delta]/TM3, Sb[1] | BDSC | FBst0006676 RRID: BDSC_6676 |
| D. melanogaster: Rac1 Rac2 Mtl mutant: y[1] w[*]; Rac1[J11] Rac2[Delta] P{w[+mW.hs]=FRT(w[hs])}2A Mtl[Delta]/TM6B, Tb[1] | BDSC | FBst0006678 RRID: BDSC_6678 |
| D. melanogaster: y1 w*; P{UASp-YFP.Rab5.S43N}Eip75B02/TM3, Sb1 | BDSC | FBst0009772 RRID: BDSC_9772 |
| D. melanogaster: y1 w*; P{UASp-YFP.Rab11.S25N}06 | BDSC | FBst0023261 RRID: BDSC_23261 |
| D. melanogaster: RNAi of tsu y1 v1; P{TRiP.HM05166}attP2 | BDSC | FBst0028955 RRID: BDSC_28955 |
| D. melanogaster: RNAi of SmB y1 v1; P{TRiP.HM05097}attP2 | BDSC | FBst0028887 RRID: BDSC_28887 |
| D. melanogaster: RNAi of SmD3 y1 sc* v1 sev21; P{TRiP.HM05226}attP2 | BDSC | FBst0030534 RRID: BDSC_30534 |
| D. melanogaster: RNAi of noi y1 sc* v1 sev21; P{TRiP.HMS00163}attP2 | BDSC | FBst0034845 RRID: BDSC_34845 |
| D. melanogaster: RNAi of gkt y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.HMJ24083}attP40 | BDSC | FBst0062874 RRID: BDSC_62874 |
| D. melanogaster: Sec23 mutant ru[1] h[1] Diap1[1] st[1] Sec23[9G] cu[1] sr[1] e[s] ca[1]/TM3, Sb[1] Ser[1] | BDSC | FBst0003094 RRID: BDSC_3094 |
| D. melanogaster: Sec24CD mutant cn[1] Sec24CD[1] bw[1] speck[1]/CyO | BDSC | FBst0003264 RRID: BDSC_3264 |
| D. melanogaster: tws mutant P{ry[+t7.2]=PZ}tws[02414] ry[506]/TM3, ry[RK] Sb[1] Ser[1] | BDSC | FBst0011568 RRID: BDSC_11568 |
| Oligonucleotides | ||
| Primers for PCR out cactin cDNA: cactin-F: 5′-TTGGGAATTCGAGCTCATGCCCAAGGAGAAATCCAAGC-3′ cactin-R: 5′-ATCCTCTAGAGGATCTACTAGTCTTATCGATTCACCGCCTGTAGCGATAGCGCTTGAA-3′ | This work | N/A |
| Primers for qRT-PCR RP49 control: rp49_F: GCTAAGCTGTCGCACAAATG rp49_R: GTTCGATCCGTAACCGATGT | This work | N/A |
| Primers for qRT-PCR Crb isoforms: Crb_comF: CGCTGCAGTTCAAGTACGAG Crb_comR: CGTCCTTGTAGCCAATGTCC Crb_RA-F: GGCTCATCGGGTTACAACTG Crb_RA-R: GTTCCGTTCATGGAGCAAGT Crb_RC-F: CCCCAAGAAAACTGCTTGAA Crb_RC-R: ATTGCAGTTGATGGGTGTCA | This work | N/A |
| Primers for qRT-PCR aPKC isoforms: aPKC_comF: TCCCGATCAGAACACTGAGG aPKC_comR: CGGGATTCTTGTTGAGGAAA aPKC_N2F: CCCAAGAACCGGCTGTACTA aPKC_N1R: ATCATCCGGGGTCAACTGTA aPKC_N4F: CGTCAGCCATCCCTTCTTTA aPKC_N3R: GTACAGCCGGTTCTTGGGTA aPKC_N6F: TTTACCCGATGGGCAAACTA aPKC_N5R: CTGGAAGATGTGTCCGTTGA aPKC_N7F: AAACGGCCACTTTTATGAGC aPKC_N8F: AGGAGATCGAGCCAGCCTAT aPKC_N10F: AACGATCCCTGCACCATATC aPKC_N9R: GCCATCGCAAGACAATCC | This work | N/A |
| Primers for qRT-PCR Rab5/7/11: rab5F: CGGCCTTTCTGACACAGACT rab5R: CCTGCGCTCCTCGATAATAC rab7F: CACGATCGGAGCTGATTTCT rab7R: TAGACAAGCACGCAGCAGTC rab11F: CCGGTGTTGGCAAAAGTAAT rab11R: TCCCAGATTTGCGCTTTAAT | This work | N/A |
| Recombinant DNA | ||
| Plasmid: pUAST-attB | https://www.flyc31.org | GenBank EF362409.1 |
| Software and algorithms | ||
| FIJI | PMID: 22743772 | https://fiji.sc/ |
| Adobe Illustrator CC | Adobe | N/A |
| Prism 8 GraphPad | GraphPad Software | https://www.graphpad.com/scientific-software/prism/ |
| Reagent or resource . | Source . | Identifier . |
|---|---|---|
| Antibodies | ||
| Alexa Fluor 568 phalloidin (1:500 dilution) | Thermo Fisher Scientific | Cat#A12380 |
| Mouse monoclonal anti-Arm (1:75 dilution) | DSHB | DSHB Cat#N2.7A1; RRID: AB_528089 |
| Rat monoclonal anti-Ecad (1:50 dilution) | DSHB | DSHB Cat#DCAD2; RRID: AB_528120 |
| Rabbit polyclonal anti-GFP (1:300 dilution) | Invitrogen | Cat#A10260; RRID: AB_2534022 |
| Rabbit polyclonal anti-mCherry (1:500 dilution) | Novus Bio- logicals | Cat#NBP2-25157 |
| Mouse monoclonal anti-Cactus (1:10 dilution) | DSHB | DSHB Cat#anti-cactus 3H12; RRID: AB_528109 |
| Mouse monoclonal anti-Dorsal (1:10 dilution) | DSHB | DSHB Cat#anti-dorsal 7A4; RRID: AB_528204 |
| Mouse monoclonal anti-FasIII (1:10 dilution) | DSHB | DSHB Cat#7G10 anti-Fasciclin III; RRID: AB_528238 |
| Mouse monoclonal anti-Dlg (1:10 dilution) | DSHB | DSHB Cat#4F3 anti-discs large; RRID: AB_528238 |
| Mouse monoclonal anti-Crb (1:10 dilution) | DSHB | DSHB Cat#cq4; RRID: AB_528181 |
| Rabbit anti-aPKC (1:200 dilution) | Santa Cruz Biotech-nology | Cat#sc-216 |
| Chemicals, peptides, and recombinant proteins | ||
| Hoechst 33342 | Sigma-Aldrich | Cat# 14533 |
| Experimental models: organisms/strains | ||
| D. melanogaster: c306-Gal4: P{w[+mW.hs]=GawB}c306, w[1118] | BDSC | FBti0003935 RRID: BDSC_3743 |
| D. melanogaster: y[1] w[*]; P{y[+t*] w[+mC]=UAS-Lifeact-GFP}VIE-260B | Hatan et al. (2011) | FBtp0064437 |
| D. melanogaster: w[1118] | BDSC | FBst0003605 RRID: BDSC_3605 |
| D. melanogaster: RNAi of Cactin: w1118; P{GD9200}v32718 | Vienna Drosophila Resource Center | FBti0098576 RRID: v32718 |
| D. melanogaster: RNAi of Cactin w1118; P{GD9200}v32719 | Vienna Drosophila Resource Center | FBti0098576 RRID: v32719 |
| D. melanogaster: RNAi of Cactin P{KK100507}VIE-260B | Vienna Drosophila Resource Center | FBti0116514 RRID: v106979 |
| D. melanogaster: P{tubP-GAL80ts}2 | BDSC | FBti0027797 RRID: BDSC_7017 |
| D. melanogaster: UAS-Cactin | This work | N/A |
| D. melanogaster: UAS-Cactin-RFP | This work | N/A |
| D. melanogaster: P{UAS-mCherry.CAAX.S}2 | BDSC | FBti0164914 RRID: BDSC_59021 |
| D. melanogaster: Fruitless-Gal4 | Borensztejn et al. (2013) | N/A |
| D. melanogaster: Upd-Gal4: P{GawB}E132, w* | BDSC | FBti0002638 RRID: BDSC_26796 |
| D. melanogaster: RNAi of white: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.HMS00017}attP2 | BDSC | FBst0033623 RRID: BDSC_33623 |
| D. melanogaster: Hsflp; AY-Gal4, UAS-GFP | D. Montell lab stock | N/A |
| D. melanogaster: w[1118]; P{10XStat92E-GFP}1 | BDSC | FBst0026197 RRID: BDSC_26197 |
| D. melanogaster: UAS-cactus: M{UAS-cact.ORF.3xHA}ZH-86Fb | FlyORF | FBti0153388 |
| D. melanogaster: w[*]; P{w[+mC]=sqh-mCherry.M}3 | BDSC | FBst0059024 RRID: BDSC_59024 |
| D. melanogaster: y1 w*; P{UASp-YFP.Rab5}02 | BDSC | FBti0100788 RRID: BDSC_24616 |
| D. melanogaster: y1 w*; P{UASp-YFP.Rab7}vig218 | BDSC | FBst0023270 RRID: BDSC_23270 |
| D. melanogaster: y1 w*; P{UASp-YFP.Rab11}sra32/TM3, Sb1 | BDSC | FBst0009790 RRID: BDSC_9790 |
| D. melanogaster: apkc mutant: y1 w67c23; P{lacW}aPKCk06403/CyO | BDSC | FBst0010622 RRID: BDSC_10622 |
| D. melanogaster: EGFP-aPKC | Chen et al. (2018) | FBti0211865 |
| D. melanogaster: UAS-GrabFP-Apical: M{lexAop-UAS-GrabFP.A.Int.mCh}ZH-86Fb | BDSC | FBti0187198 RRID: BDSC_68178 |
| D. melanogaster: UAS-GrabFP-Basal: M{lexAop-UAS-GrabFP.B.Int.mCh}ZH-86Fb | BDSC | FBst0068175 RRID: BDSC_68175 |
| D. melanogaster: UAS-Rac-FRET: P{w[+mC]=UAS-YPet-PAK-RAC-CFP.FRET}3 | BDSC | FBst0031431 RRID: BDSC_31431 |
| D. melanogaster: Rac2 mutant: Rac2[Delta] ry[506] | BDSC | FBst0006675 RRID: BDSC_6675 |
| D. melanogaster: Mtl mutant: y[1] w[*]; P{ry[+t7.2]=neoFRT}82B Mtl[Delta]/TM3, Sb[1] | BDSC | FBst0006676 RRID: BDSC_6676 |
| D. melanogaster: Rac1 Rac2 Mtl mutant: y[1] w[*]; Rac1[J11] Rac2[Delta] P{w[+mW.hs]=FRT(w[hs])}2A Mtl[Delta]/TM6B, Tb[1] | BDSC | FBst0006678 RRID: BDSC_6678 |
| D. melanogaster: y1 w*; P{UASp-YFP.Rab5.S43N}Eip75B02/TM3, Sb1 | BDSC | FBst0009772 RRID: BDSC_9772 |
| D. melanogaster: y1 w*; P{UASp-YFP.Rab11.S25N}06 | BDSC | FBst0023261 RRID: BDSC_23261 |
| D. melanogaster: RNAi of tsu y1 v1; P{TRiP.HM05166}attP2 | BDSC | FBst0028955 RRID: BDSC_28955 |
| D. melanogaster: RNAi of SmB y1 v1; P{TRiP.HM05097}attP2 | BDSC | FBst0028887 RRID: BDSC_28887 |
| D. melanogaster: RNAi of SmD3 y1 sc* v1 sev21; P{TRiP.HM05226}attP2 | BDSC | FBst0030534 RRID: BDSC_30534 |
| D. melanogaster: RNAi of noi y1 sc* v1 sev21; P{TRiP.HMS00163}attP2 | BDSC | FBst0034845 RRID: BDSC_34845 |
| D. melanogaster: RNAi of gkt y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.HMJ24083}attP40 | BDSC | FBst0062874 RRID: BDSC_62874 |
| D. melanogaster: Sec23 mutant ru[1] h[1] Diap1[1] st[1] Sec23[9G] cu[1] sr[1] e[s] ca[1]/TM3, Sb[1] Ser[1] | BDSC | FBst0003094 RRID: BDSC_3094 |
| D. melanogaster: Sec24CD mutant cn[1] Sec24CD[1] bw[1] speck[1]/CyO | BDSC | FBst0003264 RRID: BDSC_3264 |
| D. melanogaster: tws mutant P{ry[+t7.2]=PZ}tws[02414] ry[506]/TM3, ry[RK] Sb[1] Ser[1] | BDSC | FBst0011568 RRID: BDSC_11568 |
| Oligonucleotides | ||
| Primers for PCR out cactin cDNA: cactin-F: 5′-TTGGGAATTCGAGCTCATGCCCAAGGAGAAATCCAAGC-3′ cactin-R: 5′-ATCCTCTAGAGGATCTACTAGTCTTATCGATTCACCGCCTGTAGCGATAGCGCTTGAA-3′ | This work | N/A |
| Primers for qRT-PCR RP49 control: rp49_F: GCTAAGCTGTCGCACAAATG rp49_R: GTTCGATCCGTAACCGATGT | This work | N/A |
| Primers for qRT-PCR Crb isoforms: Crb_comF: CGCTGCAGTTCAAGTACGAG Crb_comR: CGTCCTTGTAGCCAATGTCC Crb_RA-F: GGCTCATCGGGTTACAACTG Crb_RA-R: GTTCCGTTCATGGAGCAAGT Crb_RC-F: CCCCAAGAAAACTGCTTGAA Crb_RC-R: ATTGCAGTTGATGGGTGTCA | This work | N/A |
| Primers for qRT-PCR aPKC isoforms: aPKC_comF: TCCCGATCAGAACACTGAGG aPKC_comR: CGGGATTCTTGTTGAGGAAA aPKC_N2F: CCCAAGAACCGGCTGTACTA aPKC_N1R: ATCATCCGGGGTCAACTGTA aPKC_N4F: CGTCAGCCATCCCTTCTTTA aPKC_N3R: GTACAGCCGGTTCTTGGGTA aPKC_N6F: TTTACCCGATGGGCAAACTA aPKC_N5R: CTGGAAGATGTGTCCGTTGA aPKC_N7F: AAACGGCCACTTTTATGAGC aPKC_N8F: AGGAGATCGAGCCAGCCTAT aPKC_N10F: AACGATCCCTGCACCATATC aPKC_N9R: GCCATCGCAAGACAATCC | This work | N/A |
| Primers for qRT-PCR Rab5/7/11: rab5F: CGGCCTTTCTGACACAGACT rab5R: CCTGCGCTCCTCGATAATAC rab7F: CACGATCGGAGCTGATTTCT rab7R: TAGACAAGCACGCAGCAGTC rab11F: CCGGTGTTGGCAAAAGTAAT rab11R: TCCCAGATTTGCGCTTTAAT | This work | N/A |
| Recombinant DNA | ||
| Plasmid: pUAST-attB | https://www.flyc31.org | GenBank EF362409.1 |
| Software and algorithms | ||
| FIJI | PMID: 22743772 | https://fiji.sc/ |
| Adobe Illustrator CC | Adobe | N/A |
| Prism 8 GraphPad | GraphPad Software | https://www.graphpad.com/scientific-software/prism/ |