Key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent, KI-mouse (Mus musculus male and female) | Q175 KI | CHDI-81003003 | N/A | Knock-in Mouse human HTT exon 1 sequence containing a ~190 CAG repeat tract |
| Genetic reagent, Transgenic (Mus musculus male and female) |
TRGL6 | Lee et al., 2019 | N/A | Transgenic with Thy1 promoter |
| Genetic reagent, Transgenic (Mus musculus male and female) cross with KI (Mus musculus male and female) |
Q175/TRGL | This study | N/A | Crossed the zQ175 KI with the TRGL6 mice |
| Sequence-based reagent | Forward primer for genotyping TRGL: 50-CTT TCC CCA CAG AAT CCA AGT CGG AAC-30 |
Lee et al., 2019 | ||
| Sequence-based reagent | Reverse primer for genotyping TRGL: 50-GCA CGA ATT CGG GCG CCG GTG GAG TGG CGG-30 |
Lee et al., 2019 | ||
| Antibody | tHTT | Cell Signaling Technology | Cat# 5656; RRID:AB_10827977 | WB: 1:1000 |
| Antibody | ntHTT mEM48 | Millipore sigma | Cat# MAB5374; RRID:AB_177645 | WB 1:200 IHC 1:50 |
| Antibody | HTT 1C2 | Millipore sigma | Cat# mab1574; RRID:AB_94263 | WB 1:1000 |
| Antibody | HTT human | Millipore sigma | Cat# mab5490; RRID:AB_2233522 | WB 1:1000 IHC 1:200 |
| Antibody | HTT MW8 | Develop Studies Hybridoma Bank, University of Iowa | Cat# MW8; RRID:AB_528297 | |
| Antibody | HTT PHP2 | CHDI/Coriell | Cat# CHDI-90001516–2 | |
| Antibody | MTOR | Cell Signaling Technology | Cat# 2983; RRID:AB_2105622 | WB: 1:1000 |
| Antibody | p-MTOR S2481 | Cell Signaling Technology | Cat# 2874 | WB 1:1000 |
| Antibody | p-MTOR S2448 | Cell Signaling Technology | Cat# 2971; RRID:AB_330970 | WB: 1:1000 |
| Antibody | p70S6K | Cell Signaling Technology | Cat# 2708; RRID:AB_390722 | WB: 1:1000 |
| Antibody | p-p70S6K S371 | Cell Signaling Technology | Cat#: 9208 | WB: 1:1000 |
| Antibody | p-p70S6K T389 | Cell Signaling Technology | Cat# 9234; RRID:AB_2269803 | WB: 1:1000 |
| Antibody | S6 ribosomal | Cell Signaling Technology | Cat# 2317; RRID:AB_2238583 | WB: 1:1000 |
| Antibody | p-S6 ribosomal S240/S244 | Cell Signaling Technology | Cat# 5364; RRID:AB_10694233 | WB: 1:1000 |
| Antibody | ULK1 | Cell Signaling Technology | Cat# 6439; RRID:AB_11178933 | WB: 1:500 |
| Antibody | p-ULK S757 | Cell Signaling Technology | Cat# 6888; RRID:AB_10829226 | WB: 1:500 |
| Antibody | p-ULK S317 | Cell Signaling Technology | Cat# 37762; RRID:AB_2922992 | WB: 1:500 |
| Antibody | ATG5 | Cell Signaling Technology | Cat# 12994; RRID:AB_2630393 | WB: 1:1000 |
| Antibody | ATG5 | Millipore sigma | Cat# abc14 | WB: 1:750 IHC 1:500 |
| Antibody | ATG14 | Cell Signaling Technology | Cat# 5504; RRID:AB_10695397 | WB: 1:1000 |
| Antibody | p-ATG14 S29 | Cell Signaling Technology | Cat# 92340; RRID:AB_2800182 | WB: 1:1000 |
| Antibody | Beclin 1 | BD Bioscience | Cat# 612113; RRID:AB_399484 | WB: 1:1000 |
| Antibody | p-Beclin 1 S30 | Cell Signaling Technology | Cat# 35955 | WB: 1:1000 |
| Antibody | VPS34 | Cell Signaling Technology | Cat# 4263 | WB 1:1000 |
| Antibody | p-VPS34 S249 | Cell Signaling Technology | Cat# 13857; RRID:AB_2798332 | WB 1:1000 |
| Antibody | TRAF6 | Cell Signaling Technology | Cat# 8028 | WB 1:1000 |
| Antibody | LC3 | Millipore sigma | Cat# abc929 | WB 1:500 IHC 1:100 |
| Antibody | LC3 | Novus Biologics | Cat# NB100-2220; RRID:AB_10003146 | WB 1:1000 IHC 1:200 |
| Antibody | p62/sqstm1 | BD Biosciences | Cat# 610832; RRID:AB_398152 | WB 1:2000 IHC 1:200 |
| Antibody | p62/SQSTM1 c-term | Progen Biotechnik | Cat# GP62-C; RRID:AB_2687531 | WB 1:1000 IHC 1:250 |
| Antibody | Ubiquitin | Dako Agilent | Cat# z0458; RRID:AB_2315524 | |
| Antibody | Ubiquitin | Abcam | Cat# ab7780; RRID:AB_306069 | |
| Antibody | LAMP1 | Develop Studies Hybridoma Bank, University of Iowa | Cat# H4A3; RRID:AB_2296838 | IHC 1:50 |
| Antibody | CTSB | Neuromics | Cat# GT15047; RRID:AB_2737184 | WB 1:5000 IHC 1:500 |
| Antibody | CTSD | In house | RU4 | WB 1:10,000 IHC 1:5000 |
| Antibody | CTSD sheep | In house | D-2–3 | WB 1:1000 IHC 1:500 |
| Antibody | DARPP32 | Abcam | Cat# ab40801; RRID:AB_731843 | WB 1:1000 IHC 1:100 |
| Antibody | NeuN | Millipore Sigma | Cat# mab377; RRID:AB_2298772 | IHC 1:400 |
| Antibody | β−actin | Millipore Sigma | Cat# A1978; RRID:AB_476692 | WB 1:10,000 |
| Antibody | Goat anti-rabbit secondary | Vector Laboratories | Cat# 68–4140 | IHC: 1:500 |
| Antibody | Goat anti-mouse secondary | Vector Laboratories | Cat# BA-9200 | IHC: 1:500 |
| Antibody | Alexa Fluor 488-conjugated goat anti-rabbit IgG | Thermo Fisher Scientific | Cat# A11034; RRID:AB_2576217 | IHC: 1:500 |
| Antibody | Alexa Fluor 568- goat anti-rabbit IgG | Thermo Fisher Scientific | Cat# A11036; RRID:AB_10563566 | IHC: 1:500 |
| Antibody | Alexa Fluor 647- goat anti-rabbit IgG | Thermo Fisher Scientific | Cat# A21245; RRID:AB_2535813 | IHC: 1:500 |
| Antibody | Alexa Fluor 405- goat anti-rabbit IgG | Thermo Fisher Scientific | Cat# A48254; RRID:AB_2890548 | IHC: 1:500 |
| Antibody | Alexa Fluor 568- goat anti-mouse IgG | Thermo Fisher Scientific | Cat# A11031; RRID:AB_144696 | IHC: 1:500 |
| Antibody | Alexa Fluor 647- goat anti-mouse IgG | Thermo Fisher Scientific | Cat# A21235; RRID:AB_2535804 | IHC: 1:500 |
| Antibody | Alexa Fluor 568- goat anti-rat IgG | Thermo Fisher Scientific | Cat# A21247; RRID:AB_141778 | IHC: 1:500 |
| Antibody | Donkey anti-Rabbit IgG HRP | Jackson ImmunoResearch | Cat# 711-035-152; RRID:AB_10015282 | WB: 1:5000 |
| Antibody | Donkey anti-Mouse IgG HRP | Jackson ImmunoResearch | Cat# 712-035-150; RRID:AB_2340638 | WB: 1:5000 |
| Antibody | Donkey anti-goat IgG HRP | Jackson ImmunoResearch | Cat# 705-035-003; RRID:AB_2340390 | WB: 1:5000 |
| Antibody | 10 nm gold anti mouse IgG | Electron Microscopy Sciences | Cat# 25129 | IEM 1:50 |
| Antibody | 10 nm gold anti rabbit IgG | Electron Microscopy Sciences | Cat# 25109 | IEM 1:50 |
| Chemical compound, drug | INK (mTOR i) | ChemScene | CAS 1224844-38-5 | Dissolved in 0.5% carboxymethyl cellulose (Sigma, Cat #5678) and 0.05% tween 80 in water |
| Chemical compound, drug | PBS | Thermo Fisher Scientific | Cat# BP339-4 | |
| Chemical compound, drug | Sodium cacodylate buffer | Electron Microscopy Sciences | Cat#11652 | 0.1 M for fixation of brains |
| Chemical compound, drug | Paraformaldehyde | Electron Microscopy Sciences | Cat#15714 | 4% for fixation of brains |
| Chemical compound, drug | 25% glutaraldehyde | Electron Microscopy Sciences | Cat# 16220 | |
| Chemical compound, drug | Uranyl acetate | Electron Microscopy Sciences | Cat# 22400–4 | EM processing |
| Chemical compound, drug | Lead citrate | Electron Microscopy Sciences | Cat#22410 | EM processing |
| Chemical compound, drug | Osmium tetroxide | Ted Pella | Cat#18465 | EM processing |
| Chemical compound, drug | Sodium metaperiodate | Sigma-Aldrich | Cat#S1878-25g | EM processing |
| Chemical compound, drug | Permount | Electron Microscopy Sciences | Cat# 17986 | Dab IHC |
| Chemical compound, drug | Flouro-gel | Electron Microscopy Sciences | Cat# 17985–10 | IF IHC |
| Chemical compound, drug | FBS | Thermo Fisher Scientific | Cat# 26140 | IHC blocking |
| Chemical compound, drug | Horse serum | Thermo Fisher Scientific | Cat# 16050 | IHC blocking |
| Chemical compound, drug | Spurr resin | Electron Microscopy Sciences | Cat# 14300 | |
| Commercial assay kit | Vectastain ABC | Vector Laboratories | Cat# PK-4000; RRID:AB_2336818 | For DAB staining |
| Commercial assay kit | DAB Peroxidase Substrate kit | Vector Laboratories | Cat# SK-4100; RRID:AB_2336382 | For DAB staining |
| Commercial assay kit | HQ silver kit | NanoProbes | Cat# 2012–45 ml | EM processing |
| Software | ImageJ | NIH | https://imagej.nih.gov/ij/ | |
| Software | Excel | Microsoft | Microsoft 365 | |
| Software | GraphPad Prism 8.0.1 | GraphPad | ||
| Other | Nitrocellulose membrane | Whatman | WB 0.2µm-pore | |
| Other | Aclar embedding film | Electron Microscopy Sciences | Cat# 50425–25 | EM processing |
| Other | 75 mesh nickel grids with carbon coating and formvar | Electron Microscopy Sciences | Cat# pi-75-ni-25 | EM processing |
| Other | EM block | Electron Microscopy Sciences | Cat# 25596 | EM processing |