KEY RESOURCES TABLE
REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
Chicken polyclonal anti-MAP2, for ICC 1:250 | Aves Labs | Cat# MAP, RRID:AB_2313549 |
Mouse monoclonal anti-Tau13, for IB 1:1000 | provided by Peter Davies (Northwell) | N/A |
Rabbit polyclonal anti-TIA1, for ICC 1:300, for IB 1:800 | Abcam | Cat# ab40693, RRID:AB_2201438 |
Goat polyclonal anti-mCherry, for ICC 1:300, for IB 1:500 | MyBioSource | Cat# MBS448057 |
Mouse monoclonal anti-TOC1, for IB 1:500 | provided by Nicholas Kanaan | N/A |
Rabbit monoclonal anti-Lamin B2, for ICC 1:500, for IB 1:1000 | Abcam | Cat# ab151735 |
Mouse monoclonal anti-Puromycin, clone 12D10, for IB 1:1000, for ICC 1: 500 | Millipore Sigma | Cat# MABE343, RRID:AB_2566826 |
Rabbit monoclonal anti-EIF2α, for IB 1:500 | Cell Signaling Technology | Cat# 5324, RRID:AB_10692650 |
Rabbit monoclonal phospho-eIF2α (Ser51) (119A11), for IB 1:500 | Cell Signaling Technology | Cat# 3597, RRID: AB_390740 |
Rabbit monoclonal anti-Cleaved Caspase 3 (Asp175) (5A1E), for ICC 1:400 | Cell Signaling Technology | Cat# 9664, RRID:AB_2070042 |
Rabbit Polyclonal anti-eIF3l, for IB 1:500 | Thermo Fisher Scientific, Invitrogen | Cat# PA531647, RRID:AB_2549120 |
Mouse monoclonal anti-TOMA2, for ICC 1:300 | provided by Rakez Kayed | N/A |
Rabbit Polyclonal anti-hnRNP H, for IB 1:500 | Bethyl Labs | Cat# A300-511A |
Rabbit Polyclonal anti-PAWR, for IB 1:500 | Thermo Fisher Scientific | Cat# 20688-1-AP, RRID:AB_10733473 |
Mouse monoclonal phospho-Tau (Thr181) antibody AT270, for ICC 1:400 | Thermo Fisher Scientific | Cat# MN1050, RRID:AB_223651 |
Mouse monoclonal phospho-Tau (S262) antibody 12E8, for ICC 1:400 | provided by Philip Dolan, Prothena | N/A |
Rabbit Polyclonal anti-hnRNP A2B1, IHC, 1:300 | Thermo Fisher Scientific | Cat# PA534939, RRID:AB_2552288 |
Rabbit Polyclonal anti-m6a | Synaptic Systems | Cat# 202 003, RRID:AB_2279214 |
Rabbit Polyclonal anti-Lbr, IHC, 1:300 | Proteintech | Cat# 12398-1-AP, RRID:AB_2138334 |
Rabbit Polyclonal anti-TDP43, IHC, 1:300 | Proteintech | Cat# 12892-1-AP, RRID:AB_2200505 |
Alexa Fluor594 Donkey Anti-Mouse IgG | Jackson ImmunoResearch | Cat#715-585-150, RRID:AB_2340854 |
Alexa Fluor488 Donkey Anti-Rabbit IgG | Jackson ImmunoResearch | Cat#711-545-152, RRID:AB_2313584 |
DyLight405 Donkey Anti-Chicken IgG | Jackson ImmunoResearch | Cat#703-475-155, RRID:AB_2340373 |
Alexa Fluor647 Donkey Anti-Chicken IgG | Jackson ImmunoResearch | Cat#703-605-155, RRID:AB_2340379 |
RFP-Trap manganese beads | Chromotek | Cat# 20 |
Bacterial and virus strains | ||
pHR-mCherry-Cry2 Lentivirus | This paper | N/A |
pHR-4R1N Tau-mCherry-Cry2 Lentivirus | This paper | N/A |
NEB Stable Competent E. coli (High Efficiency) | New England Biolabs | Cat# C3040H |
Biological samples | ||
Human temporal superior gyrus tissues (Broadman areas 41/42) | provided by Boston medical center and The Mount Sinai Hospital | N/A |
Human Ctrl/AD brain cortex of each Braak stage | Provided by Emory U. | N/A |
Chemicals, peptides, and recombinant proteins | ||
FuGENE HD transfection reagent | Promega | Cat# E2311 |
Lenti-X™ Concentrator | Takara Bio USA | Cat# 631231 |
Neurobasal medium | Thermo Fisher Scientific | Cat#21103049 |
B-27 Supplement (50x), serum free | Life Technologies | Cat#17504-044 |
DNase I Solution (1 mg/mL) | STEMCELL Technologies | Cat #07900 |
Critical commercial assays | ||
LDH kit | Promega | Cat#G1780 |
RFP-Trap Magnetic Agarose beads IP kit | ChromoTek | Cat# rtma-20 |
NuPAGE™ 3-8% Tris-Acetate Protein Gels | Invitrogen | Cat# EA03785BOX |
PLA Duolink™ In Situ Orange Starter Kit Mouse/Rabbit | Millipore Sigma | Cat# DUO92102 |
EpiQuik m6A RNA Methylation Quantification Kit | Epigentek | Cat# P-9005-96 |
Deposited data | ||
Raw Imaging Files | This study, Mendeley Data | http://dx.doi.org/10.17632/p288s6snps.1 |
mass spectrometry of mcherry IP samples | This paper | Table S1 |
Experimental models: Cell lines | ||
HEK-293T cell line | ATCC | Cat# CRL-11268 |
Tau RD P301S FRET Biosensor line | ATCC | Cat# CRL-3275 |
Experimental models: Organisms/strains | ||
C57BL/6 primary cortical neurons | This paper | |
PS19 P301S tau transgenic mice | Jackson Laboratories | stock# 008169 |
Oligonucleotides | ||
Recombinant DNA | ||
pHR-mCherry-Cry2 plasmid | This paper | N/A |
pHR-4R1N Tau-mCherry-Cry2 plasmid | This paper | N/A |
V-SVG helper plasmid | provided by Clifford P. Brangwynne lab | N/A |
PSP helper plasmid | provided by Clifford P. Brangwynne lab | N/A |
Software and algorithms | ||
Graphpad Prism 9 | https://www.graphpad.com/scientific-software/prism/ | Need to purchase license |
R package limma | Ref. Ritchie et al., 2015 | N/A |
STRING | https://string-db.org/ | N/A |
Cytoscape 3.7.2 | https://cytoscape.org/download_old_versions.html | N/A |
Fiji ImageJ | https://imagej.net/software/fiji/ | N/A |
Other | ||
490λ LED light source | This paper | N/A |
Black frame plate with high performance #1.5 cover glass | Cellvis | Cat# P24-1.5H-N |
35 mm Dish, High Precision 1.5 Coverslip, 14 mm Glass Diameter | MatTek | Cat# P35G-0.170-14-C |
Optical Power and Energy Meter | Thorlabs | Ref# PM100D |