Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Recombinant DNA reagent | pRK5-myc-mGlu7 WT | Dr. Katherine W. Roche (NIH) | ||
| Recombinant DNA reagent | pRK5-FLAG-mGlu7 WT | This study | ||
| Recombinant DNA reagent | pRK5-myc-mGlu7 Δ857 | This study | ||
| Recombinant DNA reagent | pRK5-myc-mGlu7 Δ860 | This study | ||
| Recombinant DNA reagent | pRK5-myc-mGlu7 Δ879 | This study | ||
| Recombinant DNA reagent | pRK5-myc-mGlu7 Δ893 | This study | ||
| Recombinant DNA reagent | pRK5-myc-mGlu7 CT 8K8R | This study | ||
| Recombinant DNA reagent | pRK5-myc-mGlu7 iL 4K4R | This study | ||
| Recombinant DNA reagent | pRK5-myc-mGlu7 12K12R | This study | ||
| Recombinant DNA reagent | pRK5-myc-mGlu7 iL K688/689R | This study | ||
| Recombinant DNA reagent | pRK5-myc-mGlu7 Δ857/iL 4K4R | This study | ||
| Recombinant DNA reagent | pRK5-myc-mGlu7 Δ857/IL K688/689R | This study | ||
| Recombinant DNA reagent | FLAG-ß2AR | Addgene | RRID:Addgene_14697 | |
| Recombinant DNA reagent | pRK5-HA-Ub WT | Addgene | RRID:Addgene_17608 | |
| Recombinant DNA reagent | pRK5-HA-Ub K48 | Addgene | RRID:Addgene_17605 | |
| Recombinant DNA reagent | pRK5-HA-Ub K63 | Addgene | RRID:Addgene_17606 | |
| Recombinant DNA reagent | pGFP-N3-ß-arrestin1-GFP | Dr. Katherine W. Roche (NIH) | ||
| Recombinant DNA reagent | pGFP-N3-ß-arrestin2-GFP | Dr. Katherine W. Roche (NIH) | ||
| Recombinant DNA reagent | pcDNA3-ß-arrestin1-FLAG | Addgene | RRID:Addgene_14687 | |
| Recombinant DNA reagent | pCI-HA-Nedd4 WT | Addgene | RRID:Addgene_27002 | |
| Recombinant DNA reagent | pCI-HA-Nedd4 C867S | This study | ||
| Recombinant DNA reagent | FLAG-Nedd4 WT | This study | ||
| Recombinant DNA reagent | FLAG-Nedd4 C867S | This study | ||
| Recombinant DNA reagent | FLAG-Nedd4 ΔC2 | This study | ||
| Recombinant DNA reagent | FLAG-Nedd4 ΔC2-WW1/2 | This study | ||
| Recombinant DNA reagent | FLAG-Nedd4 ΔWW3 | This study | ||
| Recombinant DNA reagent | FLAG-Nedd4 ΔWW4 | This study | ||
| Recombinant DNA reagent | FLAG-Nedd4 ΔHECT | This study | ||
| Recombinant DNA reagent | pSuper-rat Nedd4 KD | This study | ||
| Recombinant DNA reagent | pSuper-human ß-arrestin1 KD | This study | ||
| Recombinant DNA reagent | pSuper-human ß-arrestin2 KD | This study | ||
| Recombinant DNA reagent | pSuper-rat ß-arrestin1 KD | This study | ||
| Recombinant DNA reagent | pSuper-rat ß-arrestin2 KD | This study | ||
| Recombinant DNA reagent | pSuper-non-related target KD | This study | ||
| Recombinant DNA reagent | FHUGW-rat ß-arrestin1 KD | This study | ||
| Recombinant DNA reagent | FHUGW-rat ß-arrestin2 KD | This study | ||
| Recombinant DNA reagent | FHUGW-non-related target KD | This study | ||
| Recombinant DNA reagent | FHUGW-myc-mGlu7 WT | This study | ||
| Recombinant DNA reagent | FHUGW-myc-mGlu7 CT 8K8R | This study | ||
| Recombinant DNA reagent | FHUGW-myc-mGlu7 iL 4K4R | This study | ||
| Recombinant DNA reagent | FHUGW-myc-mGlu7 12K12R | This study | ||
| Recombinant DNA reagent | FHUGW-FLAG-Nedd4 WT | This study | ||
| Recombinant DNA reagent | FHUGW-FLAG-Nedd4 C867S | This study | ||
| Recombinant DNA reagent | pGEX-4T-1-mGlu7 iL1 | This study | ||
| Recombinant DNA reagent | pGEX-4T-1-mGlu7 iL2 | This study | ||
| Recombinant DNA reagent | pGEX-4T-1-mGlu7 iL3 | This study | ||
| Recombinant DNA reagent | pGEX-4T-1-mGlu7a CT | This study | ||
| Recombinant DNA reagent | pGEX-4T-1-mGlu7b CT | This study | ||
| Recombinant DNA reagent | pET28a-Nedd4 | This study | ||
| Recombinant DNA reagent | pET28b-Barr1 | This study | ||
| Recombinant DNA reagent | pET28b-Barr2 | This study | ||
| Antibody | HA (clone 16B12) | BioLegend | cat# 901501 RRID:AB_2565006 |
1:1000, western blotting (WB) |
| Antibody | c-myc (clone 9E10) | SIGMA | cat# M5546 RRID:AB_260581 |
1:1000, WB; 1:500, immunofluorescence staining (IF) |
| Antibody | FLAG (clone M2) | SIGMA | cat# F1804 RRID:AB_262044 |
1:1000, WB |
| Antibody | FLAG | SIGMA | cat# F7425 RRID:AB_439687 |
1:1000, WB |
| Antibody | mGlu7a | EMD Millipore | cat# 07–239 RRID:AB_310459 |
1:2000, WB |
| Antibody | Ub (clone FK2) | EMD Millipore | cat# 04–263 RRID:AB_612093 |
1:1000, WB |
| Antibody | Ub (clone P4D1) | Santa Cruz Biotechnology | cat# sc-8017 RRID:AB_2762364 |
1:1000, WB |
| Antibody | Nedd4 | EMD Millipore | cat# 07–049 RRID:AB_310351 |
1:10000, WB |
| Antibody | Nedd4 | R&D SYSTEMS | cat# MAB6218 RRID:AB_10920762 |
1:1000, WB |
| Antibody | Beta-arrestin 1 (clone E274) | Abcam | cat# ab32099 RRID:AB_722896 |
1:1000, WB |
| Antibody | Beta-arrestin 2 (clone C16D9) | Cell Signaling Technology | cat# 3857 RRID:AB_2258681 |
1:1000, WB |
| Antibody | GFP (clone B2) | Santa Cruz Biotechnology | cat# sc-9996 RRID:AB_627695 |
1:500, WB |
| Antibody | GFP | Thermo Fischer Scientific | cat# A11122 RRID:AB_221569 |
1:2000, WB |
| Antibody | ɑ-tubulin | SIGMA | cat# T6199 RRID:AB_477583 |
1:5000, WB |
| Antibody | PP1γ1 | EMD Millipore | cat# 07–1218 RRID:AB_1977432 |
1:500, WB |
| Antibody | phospho-ERK | Santa Cruz Biotechnology | cat# sc-7383 RRID:AB_627545 |
1:1000, WB |
| Antibody | ERK | Santa Cruz Biotechnology | cat# sc-94 RRID:AB_2140110 |
1:2000, WB |
| Antibody | phospho-JNK | Cell Signaling Technology |
cat# 4671 RRID:AB_331338 |
1:1000, WB |
| Antibody | JNK | Cell Signaling Technology | cat# 9252 RRID:AB_2250373 |
1:1000, WB |
| Antibody | Goat anti-Mouse IgG (H+L) antibody, Alexa Fluor 568 | Thermo Fischer Scientific | cat# A11031 RRID:AB_144696 |
1:500, IF |
| Antibody | Goat anti-Mouse IgG (H+L) antibody, Alexa Fluor 488 | Thermo Fischer Scientific | cat# A11029 RRID:AB_138404 |
1:500, IF |
| Chemical compound | Normal goat serum blocking solution | VECTOR LABORATORIES | cat# S-1000 RRID:AB_2336615 |
|
| Chemical compound | Paraformaldehyde | SIGMA | cat# 158127 | |
| Chemical compound | ProLong Antifade Kit | Thermo Fischer Scientific | cat# P7481 | |
| Chemical compound | L-Glutamic acid | TOCRIS | cat# 0218 | |
| Chemical compound | L-AP4 | TOCRIS | cat# 0103 | |
| Chemical compound | N-Ethylmaleimide (NEM) | SIGMA | cat# 04259 | |
| Chemical compound | MG132 | TOCRIS | cat# 1748 | |
| Chemical compound |
Leupeptin hemisulfate | TOCRIS | cat# 1167 | |
| Chemical compound | Cycloheximide | SIGMA | cat# C7698 | |
| Chemical compound | MLN7243 | CHEMIETEK | cat# CT-M7243 | |
| Chemical compound |
Protein G Sepharose 4 Fast Flow | GE Healthcare | cat# 17061801 | |
| Chemical compound | Protein A Sepharose, from Staphylococcus aureus | SIGMA | cat# P3391 | |
| Chemical compound | Glutathione Sepharose 4B GST-tagged resin | GE Healthcare | cat# 17075601 | |
| Chemical compound | EZ-Link Sulfo-NHS-SS-Biotin | Thermo Fischer Scientific | cat# 21331 | |
| Chemical compound | Pierce Streptavidin Agarose | Thermo Fischer Scientific | cat# 20347 | |
| Chemical compound | shRNA to rat Nedd4 | Schwarz et al., 2010 | Oligonucleotides | |
| Chemical compound | shRNA to rat beta-arrestin 1 | Simard et al., 2013 | Oligonucleotides | |
| Chemical compound | shRNA to rat beta-arrestin 2 | Molteni et al., 2009 | Oligonucleotides | |
| Chemical compound | shRNA to human beta-arrestin 1 | Shenoy et al., 2008 | Oligonucleotides | |
| Chemical compound | shRNA to human beta-arrestin 2 | Skånland et al., 2009 | Oligonucleotides | |
| Chemical compound | shRNA to non-related target | GenScript | Oligonucleotides | |
| Cell line (human) | HEK293T | ATCC | cat# CRL-3216 RRID:CVCL_0063 |
|
| Rat (Rattus norvegicus) | Primary cultured neurons | ORIENT BIO | RGD Cat# 734476 RRID:RGD_734476) |
SD rat |