| Antibodies |
|
| Anti GFP (chicken) |
Aves Labs, Tigard, OR |
Aves Labs Cat# GFP-1010, RRID:AB_2307313 |
| Anti-20S Proteasome Core Subunits Rabbit pAb |
Milipore, Billerica, MA |
Millipore Cat# ST1053-100UL, RRID:AB_437908 |
| Alexa Fluor® 488 AffiniPure Donkey Anti-Chicken IgY (IgG) (H + L) |
JacksonImmunoResearch Laboratories, West Grove, PA |
Jackson ImmunoResearch Labs Cat# 703-545-155, RRID:AB_2340375 |
| Donkey anti-Rabbit, Alexa 546 conjugate |
Waltham, MA |
Thermo Fisher Scientific Cat# A10040, RRID:AB_2534016 |
| Anti-PSD95 antibody (ab18258), rabbit polyclonal, |
Abcam, Cambridge, United Kingdom |
Abcam Cat# ab18258, RRID:AB_444362 |
| Anti-Ubiquitinylated proteins Antibody, clone FK2 |
Milipore, Billerica, MA |
Millipore Cat# 04-263, RRID:AB_612093 |
| Anti-PSD-95 antibody (clone K28/43), mouse monoclonal |
NeuroMab |
NeuroMab, Antibodies inc. 75-028, RRID: AB_2292909 |
| HA-Tag (C29F4) rabbit polyclonal |
Cell Signaling |
Cell Signaling, mAb #3724, RRID: AB_1549585 |
| ChromoTek RFP Monoclonal antibody, mouse monoclonal |
ChromoTek |
ChromoTek, 6g6, RRID: AB_2631395 |
| Anti-GAPDH hFAB™ rhodamine antibody, human FAB, one-step detection |
BioRad |
BioRad, #12004167 |
| IRDye® 800CW Donkey anti-Rabbit IgG Secondary antibody |
Li-COR |
Li-COR, 926-32213 RRID: AB_621848 |
| IRDye® 680LT Donkey anti-Mouse IgG Secondary antibody |
Li-COR |
Li-COR, 926-68022, RRID: AB_10715072 |
| HRP Goat Anti-Mouse IgG (H + L) |
ABclonal |
ABclonal, AS003, RRID: AB_2769851 |
| HRP Goat Anti-Rabbit IgG (H + L) |
ABclonal |
ABclonal, AS014, RRID: AB_2769854 |
| GFP-Trap® Magnetic Particles M-270, alpaca monoclonal nanobody |
ChromoTek |
ChromoTek, gtma, RRID: AB_2631358 |
|
| Bacterial and virus strains |
|
| One Shot™ TOP10 Chemically Competent E. coli
|
Thermo Fisher |
C404006 |
|
| Chemicals, peptides, and recombinant proteins |
|
| (S)-3,5-DHPG |
Tocris |
Cat. No. 0805 |
| B-27™ Supplement (50X), serum free |
Thermo Fisher Scientific |
Cat. No. 17504044 |
| Neurobasal Medium |
Thermo Fisher Scientific |
Cat. No. 21103049 |
| DMEM, high glucose |
Thermo Fisher Scientific |
Cat. No. 11965092 |
| Hibernate E low fluorescence |
BrainBits |
HELF |
| Poly-D-lysine hydrobromide |
Millipore |
P6407-5MG |
| Corning™ Donor Horse Serum, U.S. Sourced |
Fisher Scientific |
Cat. No. MT35030CV |
| TERGITOL |
Sigma-Aldrich |
CAS Number 127087-87-0 |
| Triton™ X-100 Surfactant |
Sigma-Aldrich |
CAS Number 9002-93-1 |
| Paraformaldehyde |
Sigma-Aldrich |
158127 |
| HLI373 |
Tocris |
Cat. No. 3503 |
| PR-619 |
Sigma-Aldrich |
SML0430 |
|
| Critical commercial assays |
|
| Duolink™ In Situ Red Starter Kit Mouse/Rabbit |
Millipore Sigma, St. Louis, MO |
DUO92101 |
|
| Experimental models: Cell lines |
|
| Neuro-2a |
Sigma Aldrich, St. Louis, MO |
ECACC Cat# 89121404, RRID: CVCL_0470 |
| SY-SH5Y cell line, ATCC |
ATCC, Manassas, Virginia |
ATCC Cat# CRL-2266, RRID: CVCL_0019 |
|
| Experimental models: Organisms/strains |
|
| C57BL/6J- |
The Jackson Laboratory, Bar Harbor, Maine |
IMSR Cat# JAX: 000664, RRID:IMSR_JAX: 000664 |
| B6.129P2-Fmr1 tm1Cgr/J |
The Jackson Laboratory, Bar Harbor, Maine |
IMSR Cat# JAX: 003025, RRID:IMSR_JAX: 003025 |
|
| Oligonucleotides |
|
| Primers for subcloning of plasmids used in this study, see method details, constructs subsection |
This paper |
N/A |
|
| Recombinant DNA |
|
| pUbFC-YN173-Ub |
Fang and Kerppola, 2004
|
N/A |
| pBiFC-JunCC155 |
Fang and Kerppola, 2004
|
N/A |
| Venus-1-155-Ubiquitin plasmid |
This paper |
N/A |
| Venus-1-155-Ubiquitin G76A mutant |
This paper |
N/A |
| Venus-1-155-ΔUbiq |
This paper |
N/A |
| Venus (156-239)-PSD-95 plasmid |
This paper |
N/A |
| Venus (156-239)-PSD-95 lysine mutant |
This paper |
N/A |
| Venus-PSD-95 |
Ifrim et al., 2015
|
N/A |
| Venus-PSD-95 lysine mutant |
This paper |
N/A |
| UbFC-GluA1-Venus(156-239) |
This paper |
N/A |
| Venus(156-239)-FMRP-3UTR |
This paper |
N/A |
| His-Venus |
Ifrim et al., 2015
|
N/A |
| pcDNA 3.1 (–) |
ThermoFisher |
V79520 |
| pmScarlet-i_C1 |
Addgene plasmid # 85044 |
RRID: Addgene_85044 |
|
| Software and algorithms |
|
| ImageJ |
(Schneider et al., 2012) |
https://imagej.nih.gov/ij/
|
| OCTANE ImageJ Plugin |
(Tatavarty et al., 2012) |
https://health.uconn.edu/yu-lab/software
|
|
| Other |
|
| Lipofectamine 2000 Transfection reagent |
Thermo Fisher Scientific |
11668019 |
| QuickChange II XL Site-Directed Mutagenesis Kit |
Agilent Technologies, La Jolla, CA |
200521 |
| ProLong™ Diamond Antifade Mountant |
Thermo Fisher |
P36970 |
| PEI MAX 40K |
Avantor 75800-188, Polysciences Inc |
CAS Number: 49553-93-7 |
| cOmplete™, EDTA-free Protease Inhibitor Cocktail |
Roche |
04693116001 |
| Pierce BCA Protein Assay Kit |
Thermo Fisher Scientific |
Cat # 23225 |
| GFP-Trap Magnetic Beads |
Chromotek |
gtma |
| Tanon™ High-sig ECL Western Blotting Substrate |
(ABclonal) |
Cat# 180-5001 |
