Key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Drosophila melanogaster) | Nephrin (Sns) |
Flybase | FLYB: FBgn0024189 | For simplicity we use the human name |
| Gene (Drosophila melanogaster) | Neph1 (Kirre) |
Flybase | FLYB: FBgn0028369 | For simplicity we use the human name |
| Gene (Drosophila melanogaster) | Rab5 | Flybase | FLYB: FBgn0014010 | |
| Gene (Drosophila melanogaster) | Rab7 | Flybase | FLYB: FBgn0015795 | |
| Gene (Drosophila melanogaster) | Rab11 | Flybase | FLYB: FBgn0015790 | Transgenic animals |
| Gene (Drosophila melanogaster) | Shibire (Shi) | Flybase | FLYB: FBgn0003392 | Transgenic animals |
| Gene (Drosophila melanogaster) | Flotillin 2 (Flo2) | Flybase | FLYB: FBgn0264078 | Transgenic animals |
| Strain, strain background (Drosophila melanogaster) | Nephrin-RNAi (Sns-RNAi) |
VDRC | VDRC #109442 |
Transgenic animals |
| Strain, strain background (Drosophila melanogaster) | Nephrin-RNAi-2 (Sns-RNAi-2) |
BDSC | BDSC #64872 |
Transgenic animals |
| Strain, strain background (Drosophila melanogaster) | UAS-Rab5-RNAi | BDSC | BDSC #34832 |
Transgenic animals |
| Strain, strain background (Drosophila melanogaster) | UAS-Rab5S43N | BDSC | BDSC #42704 |
Transgenic animals, dominant negative variant |
| Strain, strain background (Drosophila melanogaster) | UAS-YFP-Rab5Q88L | BDSC | BDSC #9774 |
Transgenic animals, constitutively active variant |
| Strain, strain background (Drosophila melanogaster) | UAS-Rab7-RNAi | BDSC | BDSC #27051 |
Transgenic animals |
| Strain, strain background (Drosophila melanogaster) | UAS-YFP-Rab7T22N | BDSC | BDSC #9778 |
Transgenic animals, dominant negative variant |
| Strain, strain background (Drosophila melanogaster) | UAS-Rab11-RNAi | BDSC | BDSC #42709 |
Transgenic animals |
| Strain, strain background (Drosophila melanogaster) | UAS-flo2-RNAi | BDSC | BDSC #40833 |
Transgenic animals |
| Strain, strain background (Drosophila melanogaster) | UAS-flo2-RNAi-2 | VDRC | VDRC #330316 |
Transgenic animals |
| Strain, strain background (Drosophila melanogaster) | Shibirets | BDSC | BDSC #2248 |
Temperature-sensitive allele |
| Strain, strain background (Drosophila melanogaster) | Dorothy-GAL4 | BDSC | BDSC #6903 |
Transgenic, GAL4-dependent expression in nephrocytes |
| Strain, strain background (Drosophila melanogaster) | prospero-GAL4 | Weavers et al., 2009 (PubMed-ID: 18971929) | Promoter derived from: FLYB: FBgn0004595 |
Transgenic, GAL4-dependent expression in nephrocytes |
| Strain, strain background (Drosophila melanogaster) | UAS-GFP-RNAi | BDSC | BDSC #41553 |
Transgenic animals, control-RNAi |
| Strain, strain background (Drosophila melanogaster) | nephrin-GFP | This work | Edited gene: FLYB: FBgn0024189 |
Insertion of GFP into the c-terminus of sns, (genomic) |
| Strain, strain background (Drosophila melanogaster) | Myc-nephrin | This work | Edited gene: FLYB: FBgn0024189 |
Insertion of Myc into exon 2 of sns (genomic) |
| Antibody | anti-Sns (nephrin, rabbit polyclonal) |
Bour et al., 2000 (PubMed-ID: 10859168) | Target: FLYB: FBgn0024189 |
1:300 for IF |
| Antibody | anti-Kirre (Neph1, guinea pig, polyclonal) | Galletta et al., 2004 (PubMed-ID: 15511638) | Target: FLYB: FBgn0028369 |
1:200 for IF |
| Antibody | anti-Rab5 (rabbit, polyclonal) | Abcam | ab18211 | 1:200 for IF |
| Antibody | anti-Rab7 (mouse, monoclonal) |
DSHB | Rab7 | 1:100 for IF |
| Antibody | anti-Myc (mouse, monoclonal) |
DSHB | 9E10 | 1:100 for IF |
| Antibody | anti-Myc (mouse, monoclonal) |
Santa Cruz Biotechnology | sc-40 | 1:100 for IF |
| Antibody | anti-RAB11 (rabbit, monoclonal) |
Cell Signaling Technology | 5589S | 1:100 for IF |
| Antibody | Alexa Fluor 488 anti-rabbit, (donkey, polyclonal) |
Thermofisher | #A-21206 | 1:200 for IF, secondary antibody |
| Antibody | Alexa Fluor 488 anti-mouse (donkey, polyclonal) |
Thermofisher | #A32766 | 1:200 for IF, secondary antibody |
| Antibody | Alexa Fluor 568 anti-rabbit (donkey, polyclonal) |
Thermofisher | #A10042 | 1:200 for IF, secondary antibody |
| Antibody | Alexa Fluor 568 anti-mouse (donkey, polyclonal) |
Thermofisher | #A10037 | 1:200 for IF, secondary antibody |
| Antibody | Alexa Fluor 568 anti-guinea pig (goat, polyclonal) |
Thermofisher | #11075 | 1:200 for IF, secondary antibody |
| Commercial assay or kit | In Situ Cell Death Detection Kit | Sigma/Roche | 11684795910 | TUNEL labeling |
| Chemical compound, drug | FITC-albumin | Sigma/Merck | A9771 | Final conc.: 0.2 mg/ml |
| Chemical compound, drug | Texas-Red-Dextran | Thermofisher | D1863 | Final conc.: 0.2 mg/ml |
| Chemical compound, drug | Texas-Red-Avidin | Thermofisher | A2348 | Final conc.: 0.2 mg/ml |
| Chemical compound, drug | Alexa 488 wheat germ agglutinin | Thermofisher | W11261 | Final conc.: 0.2 mg/ml |
| Chemical compound | Roti-Mount | Carl Roth | HP19.1 | For mounting |
| Chemical compound, drug | Hoechst 33342 | Thermofisher | H1399 | 1:1000 for IF |
| Chemical compound, drug | Bafilomycin | Invivogen | tlrl-baf1 | Final conc.: 0.1 µM |
| Chemical compound, drug | Methyl-β-cyclodextrin | Sigma-Aldrich/Merck | 332615 | Final conc.: 10 mM |
| Chemical compound,drug | Low-melting-agarose | Carl Roth | #6351.5 | Final use: 1% agarose |
| Chemical compound, drug | Schneider’s insect medium | Sigma-Aldrich/Merck | #S0146 | |
| Chemical compound, drug | Triton X-100 | Sigma-Aldrich/Merck | #9036-19-5 | Final conc.: 0.1% in PBS |
| Software, algorithm | GraphPad Prism | GraphPad Inc | GraphPad Prism 9.3.1 | |
| Software, algorithm | Fiji/ImageJ | Open source | ImageJ 2.1.0/1.53c | |
| Software, algorithm | GIMP | Open source | GIMP 2.10 |