Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Homo sapiens) | EPS15 | UniProt | P42566 | Epidermal growth factor receptor substrate 15 |
| Gene (H. sapiens) | EPS15L1;EPS15R | UniProt | Q9UBC2 | Epidermal growth factor receptor substrate 15R |
| Gene (H. sapiens) | ITSN1 | UniProt | Q15811 | Intersectin-1 |
| Gene (H. sapiens) | FCHO1; KIAA0290 | UniProt | O14526 | FCH domain only protein 1 |
| Gene (H. sapiens) | FCHO2 | UniProt | Q0JRZ9 | FCH domain only protein 2 |
| Strain, strain background (Escherichia coli) | XL1-Blue | Agilent Technologies | #200236 | |
| Strain, strain background (E. coli) | BL21 Codon Plus RIL | Agilent Technologies | #230240 | |
| Strain, strain background (E. coli) | BL21 (DE3) Codon Plus RIPL |
Agilent Technologies | #230280 | |
| Cell line (H. sapiens) (female) | HeLa SS6 | Elbashir et al., 2001 | Human cervical carcinoma cell line | |
| Cell line (H. sapiens) (female) |
HeLa SS6 clone #64/1.E | Umasankar et al., 2014 | Dual FCHO1- andFCHO2-gene disrupted (null) HeLa SS6 cells | |
| Cell line (H. sapiens) (female) | HeLa SS6 clone #64/1.E #40 (clone #40) | this paper | Triple FCHO1, FCHO2- and EPS15L1-gene disrupted (null) HeLa SS6 cells | |
| Antibody | anti-β-actin; mouse clone mAb C4 | Santa Cruz Biotechnology | sc-47779 | 1:10,000 for immunoblotting |
| Antibody | anti-AP-2 α subunit mouse mAb C-8 |
Santa Cruz Biotechnology | sc-17771 | 1:500 for immunoblotting |
| Antibody | anti-AP-2 α subunit; mouse clone mAb AP.6 | F Brodsky Laboratory | PMID: 2574457 | 1:1,000-1:2,000 for immunofluorescence |
| Antibody | anti-ARH rabbit antigen affinity purified pAb | L Traub Laboratory | PMID: 12451172 | 1:2,000 for immunoblotting |
| Antibody | anti-AP-1/2 β1/β2 subunit; rabbit antigen affinity purified pAb GD/1 | L Traub Laboratory | PMID: 7876268 | 1:2,500 for immunoblotting |
| Antibody | anti-CALM; goat affinity purified pAb C-18 | Santa Cruz Biotechnology | sc-6433 | 1:500 for immunoblotting |
| Antibody | anti-clathrin heavy chain; mouse clone mAb TD.1 | F Brodsky Laboratory | PMID: 1547490 | 1:5,000 for immunoblotting |
| Antibody | anti-Dab2 rabbit antigen affinity purified pAb | L Traub Laboratory | PMID: 12234931 | 1:2,500 for immunoblotting |
| Antibody | anti-Disabled-2/p96; mouse mAb 52/p96 | BD Transduction Laboratories | 610464 | 1:1,000 for immunoblotting |
| Antibody | anti-Eps15; rabbit affinity purified pAb C-20 | Santa Cruz Biotechnology | sc-534 | 1:500 for immunoblotting |
| Antibody | anti-Eps15; rabbit antigen affinity purified pAb | E Ungewickell Laboratory | PMID: 12960147 | 1:500-1:2,000 for immunofluorescence |
| Antibody | anti-Eps15R; rabbit mAb ab53006 | AbCAM | EP1147Y | 1:5,000 for immunoblotting |
| Antibody | anti-epsin 1; rabbit antigen affinity piurified pAb | L Traub Laboratory | PMID: 10692452 | 1:1,000 for immunoblotting |
| Antibody | anti-FCHO1; rabbit antigen affinity purified pAb |
L Traub Laboratory | 1:2,500 for immunoblotting | |
| Antibody | anti-FCHO2; rabbit antigen affinity purified pAb | Novus Biologicals | NBP2-32694 | 1:700 for immunoblotting |
| Antibody | anti-GAPDH; mouse mAb | USP Bio | Y1041 | 1:3,500 for immunoblotting |
| Antibody | anti-GFP; antigen affinity purified pAb | P Hanson Laboratory | 1:1,000 for immunoblotting | |
| Antibody | anti-GFP-Booster; llama VHH Nb | Chromotek | gba488-100 | 1:1,000-1:2000 for immunofluorescence |
| Antibody | anti-Hrb;RIP;Rab; goat affinity purified pAb | Santa Cruz Biotechnology | sc-1424 | 1:1,000 for immunoblotting |
| Antibody | anti-Hrb; AGFG1; mouse clone mAb H-2 | Santa Cruz Biotechnology | sc-166651 | 1:1,000-1:2,000 for immunofluorescence |
| Antibody | anti-intersectin 1; mouse clone mAb 29 | BD Transduction Laboratories | 611574 | 1:250 for immunoblotting |
| Antibody | anti-LAMP-1; mouse clone mAb 1D4B | Developmental Studies Hybridoma Bank | AB_528127 | 1:1,000-1:2,000 for immunofluorescence |
| Antibody | anti-mNG; mouse clone mAb 32F6 | Chromotek | 32f6-100 | 1:1,000 for immunoblotting |
| Sequence -based reagent | pGEX-5X-1 | Pharmacia | ||
| Sequence-based reagent | pGEX-4T-1 | Pharmacia | ||
| Sequence -based reagent | pMW172 | Michael Way | ||
| Sequence -based reagent | pMECS | NSF/VIB | ||
| Sequence-based reagent | pHEN6c | NSF/VIB | ||
| Sequence -based reagent | pmNG-C1 | this paper | mNG coding region replacing GFP in pEGFP-C1 vector | |
| Sequence-based reagent | pEGFP-FYVE2 | this paper | insertion of Nb E_142 following FYVE2 tandem | |
| Sequence- based reagent |
pEGFP-FYVE2-Nb E_142 | this paper | introduction of 3maj destabilizing mutations into Nb E_142 | |
| Sequence -based reagent | pEGFP-FYVE2-Nb dE_142 | this paper | ||
| Sequence -based reagent | piRFP-N1-Akap1-FRB | |||
| Sequence -based reagent | piRFP-N1-Akap1-Nb E_142 | this paper | replacement of control FRB module with Nb E_142 |
|
| Sequence -based reagent | pEGFP-FCHO1 (1–609) | Umasankar et al., 2014 | ||
| Sequence-based reagent | PCR primer sets | this paper | see Table 1 for complete PCR primer set details | |
| Peptide, recombinant protein |
bovine serum albumin (BSA) | Pierce | 23209 | |
| Peptide, recombinant protein |
glutathione | Sigma-Aldrich | G4251-1G | |
| Commercial assay or kit | QuikChange site-directed mutageneisis kit |
Agilent Technologies | 200523 | |
| Chemical compound, drug |
Glutathione- Spharose 4B |
GE Healthscience | 17075605 | |
| Chemical compound, drug |
Ni-NTA-agarose | MC Labs | NINTA-300 | |
| Chemical compound, drug | Normal goat serum | Sigma-Aldrich | 46767 | |
| Chemical compound, drug | Alexa Fluor 568-transferrin (H. sapiens) | Invitrogen; ThermoFisher Scientific | T23365 | |
| Chemical compound, drug |
Saponin | Sigma-Aldrich | S7900 | |
| Chemical compound, drug | Lipofectamine 2000 | Invitrogen;ThermoFisher Scientific | 11668027 | |
| Software, algorithm |
FV1-ASW 4.1 software | Olympus | ||
| Software, algorithm | ImageJ | Rueden et al., 2017 | ||
| Software, algorithm | Fiji | Schindelin et al., 2012 | ||
| Software, algorithm | NIS Elements 5.11.01 analysis package | Nikon |