REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
Rabbit monoclonal anti-eIF2α (phosphorylated, Ser51) | Abcam | Cat#ab32157; RRID:AB_732117 |
Rabbit polyclonal anti-ULK1 | Abcam | Cat#ab240916 |
Mouse monoclonal anti-G3BP1 | BD Biosciences | Cat#611126; RRID:AB_398437 |
Mouse monoclonal anti-TIAR | BD Biosciences | Cat#610352; RRID:AB_397742 |
Rabbit polyclonal anti-IFNAR1 | Bethyl | Cat#A304-290A; RRID:AB_2620486 |
Rabbit polyclonal anti-OPTN | Bethyl | Cat#A301-829A; RRID:AB_1264331 |
Rabbit polyclonal anti-TBK1 | Bethyl | Cat#A300-093A; RRID:AB_2303002 |
Rabbit polyclonal anti-cleaved caspase 3 | Cell Signaling | Cat#9661S; RRID:AB_2341188 |
Rabbit monoclonal anti-eIF2α (total) | Cell Signaling | Cat#D7D3; RRID:AB_10692650 |
Mouse monoclonal anti-puromycin (clone 12D10) | Merck Millipore | Cat#MABE343; RRID:AB_2566826 |
Rabbit polyclonal anti-FUS | Proteintech | Cat#11570-1-AP; RRID: AB_2247082 |
Rabbit polyclonal anti-Nup107 | Proteintech | Cat#19217-1-AP; RRID:AB_10597702 |
Rabbit polyclonal anti-Nup98-Nup96 | Proteintech | Cat#12329-1-AP; RRID:AB_10973678 |
Rabbit polyclonal anti-YBX1 | Proteintech | Cat#20339-1-AP; RRID:AB_10665424 |
Rabbit polyclonal anti-ATXN2 | Proteintech | Cat#21776-1-AP; RRID:AB_10858483 |
Rabbit polyclonal anti-KPNA2 | Proteintech | Cat#10819-1-AP; RRID:AB_2265526 |
Rabbit polyclonal anti-TNPO1 | Proteintech | Cat#20679-1-AP; RRID:AB_10694291 |
Mouse monoclonal anti-FUS | Santa Cruz | Cat#sc-47711; RRID:AB_2105208 |
Rabbit polyclonal anti-βIII-tubulin (Tuj) | Sigma-Aldrich | Cat#T2200; RRID:AB_262133 |
Mouse monoclonal anti-β-actin | Sigma-Aldrich | Cat#A5441; RRID:AB_476744 |
Bacterial and Virus Strains | ||
NEB 5-alpha Competent E. coli | New England Biolabs | Cat#C2987 |
Respiratory syncytial virus (RSV) wild-type strain A2 | Openshaw Lab 18287232 (first used in this study) | N/A |
Biological Samples | ||
Spinal cord tissue from ALS patients and healthy individuals (frozen) | Sheffield Brain Tissue Bank | Request No.15-011 |
Spinal cord tissue from ALS patients and healthy individuals (paraffin blocks) | MRC London Neurodegenerative Diseases Brain Bank | Request No.1470 |
Chemicals, Peptides, and Recombinant Proteins | ||
MG132 (proteasome inhibitor) | Calbiochem | Cat#474790 |
Pifithrin-μ (HSP70 inhibitor) | Enzo Life Sciences | Cat#BML-AP503 |
Polyinosinic–polycytidylic acid potassium salt, poly(I:C) | Sigma-Aldrich | Cat#P9582 |
Sodium arsenite | Sigma-Aldrich | Cat#35000 |
Dithiothreitol (DTT) | Sigma-Aldrich | Cat#43815 |
Interferon beta-1a | Sigma-Aldrich | Cat#IF014 |
Cycloheximide | Sigma-Aldrich | Cat#C7698 |
Puromycin dihydrochloride | Sigma-Aldrich | Cat#P8833 |
5,6-Dichlorobenzimidazole 1-β-D-ribofuranoside (DRB) | Sigma-Aldrich | Cat#D1916 |
Actinomycin D | Sigma-Aldrich | Cat#A1410 |
Critical Commercial Assays | ||
GenElute™ Total RNA Purification Kit | Sigma-Aldrich | Cat#RNB 100 |
RNase free DNase set | QIAGEN | Cat#79254 |
USB® Poly(A) Tail-Length Assay Kit | Thermo Scientific | Cat#764551KT |
FuGENE® HD transfection reagent | Promega | Cat#E2311 |
DreamTaq HS polymerase | Thermo Scientific | Cat#EP2702 |
SuperScript IV Reverse Transcriptase | Thermo Scientific | Cat# 18090010 |
Lipofectamine 2000 transfection reagent | Thermo Scientific | Cat#11668027 |
Experimental Models: Cell Lines | ||
Human: SH-SY5Y neuroblastoma cells (ATCC) | Sigma-Aldrich | Cat#94030304 |
Human: SH-SY5Y FUS line: ΔNLS1_ho (homozygous) | An et al., 2019 | N/A |
Human: SH-SY5Y FUS line: ΔNLS4_ho (homozygous) | An et al., 2019 | N/A |
Human: SH-SY5Y FUS line: ΔNLS7_ho (homozygous) | An et al., 2019 | N/A |
Human: SH-SY5Y FUS line: ΔNLS2_het (heterozygous, FUS granule positive) | An et al., 2019 | N/A |
Human: SH-SY5Y FUS line: ΔNLS8_het (heterozygous) | An et al., 2019 | N/A |
Human: SH-SY5Y FUS line: ΔNLS10_het (heterozygous) | An et al., 2019 | N/A |
Human: SH-SY5Y FUS line: ΔNLS11_het (heterozygous, FUS granule positive) | An et al., 2019 | N/A |
Human: SH-SY5Y FUS line: FUS knockout line | An et al., 2019 | N/A |
Human: primary fibroblasts (healthy control and bearing FUS P525L mutation) | Lo Bello et al., 2017 | N/A |
Human: Day 40 motor neurons derived from hES H9 cells | Shelkovnikova et al., 2017 | N/A |
Oligonucleotides | ||
Primers for qRT-PCR, see Table S1 | An et al., 2019 and this paper | N/A |
Primer: Poly(A) tail assay for FUS, P1: 5′-GTCCAGCCCA TGTGAGACTT-3′ |
This paper | N/A |
Primer: Poly(A) tail assay for FUS, P2: 5′-AACCTCCAGC ATAAAAGGGCT-3′ |
This paper | N/A |
siRNA, targeting human STAT1: MISSION® esiRNA | Sigma-Aldrich | Cat#EHU010121 |
siRNA, scrambled: AllStars Negative Control siRNA | QIAGEN | Cat#SI03650318 |
Recombinant DNA | ||
Plasmid: FUS R522G GFP | Shelkovnikova et al., 2014a | N/A |
Software and Algorithms | ||
CellF software | Olympus | N/A |
Zen software | Zeiss | https://www.zeiss.com/microscopy/int/products/microscope-software/zen.html |
ImageJ | Schneider et al., 2012 | https://imagej.nih.gov/ij/ |
Adobe Photoshop CS3 | Adobe Inc. | https://www.adobe.com/uk/products/photoshop.html |
GraphPad Prism 6 | GraphPad Software, Inc. | https://www.graphpad.com/scientific-software/prism/ |
Leica Application Suite AF software | Leica Microsystems | https://www.leica-microsystems.com/products/microscope-software/p/leica-application-suite/ |
Other | ||
BX61 microscope equipped with F-View II camera | Olympus | https://www.olympus-ims.com/en/microscope/bx61-2/ |
LSM880 microscope | Zeiss | https://www.zeiss.com/microscopy/int/products/confocal-microscopes.html |
EVOS XL Core system | Thermo Scientific | Cat#AMEX1100 |
Leica TCS SP2 MP confocal microscope | Leica Microsystems | https://www.leica-microsystems.com/products/confocal-microscopes/p/leica-tcs-sp2/ |
StepOnePlus™ Real-Time PCR System | Thermo Scientific | Cat#4376600 |