KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| rabbit anti-Rab5 | Abcam | ab31261, RRID: AB_882240 |
| rabbit anti-Rab7 | Gift from Patrick Dolph [53] | N/A |
| mouse anti-Rab11 | BD Transduction Laboratories | Clone 47, RRID: AB_397983 |
| rabbit anti-Syt1 | Developmental Studies Hybridoma Bank (DSHB) | 3H2 2D7, RRID: AB_528483 |
| rabbit anti-GABARAP+GABARAPL1+GABARAPL2 (ATG8) | Abcam | ab109364, RRID: AB_10861928 |
| guinea pig anti-Spin | Gift from Sean Sweeney [37] | N/A |
| rat anti-Sky | Gift from Patrik Verstreken [12] | N/A |
| mouse anti-CSP | DSHB | DCSP-2 (6D6), RRID: AB_528183 |
| rabbit anti-Rab3 | Gift from Aaron DiAntonio [54] | N/A |
| mouse anti-Syx1a | DSHB | 8C3, RRID: AB_528484 |
| guinea pig anti-V100 | [55] | N/A |
| rabbit anti-n-Syb | [55] | N/A |
| rat anti-n-Syb (R29) | Gift from Hugo Bellen [56] | N/A |
| mouse anti-Chaoptin | DSHB | 24B10, RRID: AB_528161 |
| Cy5 AffiniPure Goat Anti-Rabbit IgG (H+L) | Jackson Immunoresearch | 111-175-144, RRID: AB_2338013 |
| Cy5 AffiniPure Goat Anti-Mouse IgG (H+L) | Jackson Immunoresearch | 115-175-146, RRID: AB_2338713 |
| Cy5 AffiniPure Goat Anti-Rat IgG (H+L) | Jackson Immunoresearch | 112-175-143, RRID: AB_2338263 |
| Alexa Fluor 647 AffiniPuree Goat Anti-Guinea Pig IgG (H+L) | Jackson Immunoresearch | 106-605-003, RRID: AB_2337446 |
| Cy3 AffiniPure Goat Anti-Rabbit IgG (H+L) | Jackson Immunoresearch | 111-165-144, RRID: AB_2338006 |
| Alexa Fluor 488 AffiniPure Goat Anti-Rabbit IgG (H+L) | Jackson Immunoresearch | 111-545-144, RRID: AB_2338052 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| TOTO3 | Thermo Fisher | T3604 |
| Vectashield | Vector Laboratories | H-1000 |
| PBS | GIBCO | 70011-36 |
| Formaldehyde | Merck KGaA | 1.03999.1000 |
| Triton X-100 | Sigma-Aldrich | T8787 |
| Schneider’s Drosophila Medium [+] L-Glutamine | GIBCO | 21720-024 |
| Agarose, low gelling temperature | Sigma-Aldrich | A9045-10G |
| Human insulin recombinant zinc | GIBCO | 12585014 |
| Penicillin/Streptomycin | GIBCO | 15140122 |
| ES Cell FBS | GIBCO | 16141-061 |
| 20-Hydroxyecdysone | Sigma-Aldrich | 5289-74-7 |
| SilGard and Silicone Elastomer Kit | Dow Corning | 184 |
| Sodium Chloride | Merck KGaA | 1.06404.1000 |
| Experimental Models: Organisms/Strains | ||
| Drosophila, UAS-myr-mCherry-pHluorin in VK00002 | This paper | N/A |
| Drosophila, UAS-Syt1-mCherry-pHluorin in VK00002 | This paper | N/A |
| Drosophila, UAS-mCherry-pHluorin-Syt1 in VK00002 | This paper | N/A |
| Drosophila, UAS-CP1-mCherry-pHluorin in VK00002 | This paper | N/A |
| Drosophila, GMR-Gal4 | Bloomington Drosophila Stock Center (BDSC) | 9146 |
| Drosophila, ppk-Gal4 | BDSC | 32078 |
| Drosophila, ey3.5flp | BDSC | 35542 |
| Drosophila, FRT82b,tub-Gal80 | BDSC | 5135 |
| Drosophila, FRT80b,tub-Gal80 | BDSC | 5191 |
| Drosophila, nsybF33B | [57] | N/A |
| Drosophila, rab7gal4-knock-in | [17] | N/A |
| Drosophila, v1004 | [55] | N/A |
| Drosophila, sytAD4 | [58] | N/A |
| Drosophila, FRT40A,cl | BDSC | 5622 |
| Drosophila, UAS-Atg8-mCherry | BDSC | 37749 |
| Drosophila, UAS-Syt1-GFP | BDSC | 6926 |
| Drosophila, GMR-myr-GFP | BDSC | 7112 |
| Drosophila, GMR-myr-Tomato | Gift from S.Lawrence Zipursky | N/A |
| Drosophila, carΔ146,FRT19A | Gift from Helmut Kramer | N/A |
| Drosophila, UAS-V100 | [55] | N/A |
| Drosophila, UAS-n-Syb | [22] | N/A |
| Drosophila, UAS-Rab7 | Hiesinger lab | N/A |
| Drosophila, UAS-n-Syb-RNAi | VDRC | 104531 |
| Recombinant DNA | ||
| mCherry-SEpHluorin | Addgene | 32001 |
| pUASTattB | [59] | Flybase ID: FBmc0003002 |
| Software and Algorithms | ||
| ImageJ | National Institutes of Health (NIH) | N/A |
| Imaris | Bitplane, Switzerland | N/A |
| GraphPad Prism | GraphPad Software, La Jolla, USA | N/A |
| AutoQuant 3x | Media Cybernetics | N/A |
| Clampfit | Axon Instruments | N/A |
| Clampex | Axon Instruments | N/A |
| Python | Python Software Foundation | N/A |
| Amira | FEI-Thermo Fisher Scientific | N/A |