REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
pan-Keratin | Cell Signaling Technology | Cat# 4545; RRID:AB_490860 |
Emerin | Cell Signaling Technology | Cat# 5430; RRID:AB_10691714 |
ERp57 | Cell Signaling Technology | Cat# 2881; RRID:AB_2160840 |
B23-NPM | Cell Signaling Technology | Cat# 3542; RRID:AB_2155178 |
Nup98 | Cell Signaling Technology | Cat# 2598; RRID:AB_2267700 |
β-actin | Cell Signaling Technology | Cat# 3700; RRID:AB_2242334 |
eEF2k | Cell Signaling Technology | Cat# 3691; RRID:AB_2097313 |
eIF4E | Cell Signaling Technology | Cat# 9742; RRID:AB_823488 |
4E-BP1 | Cell Signaling Technology | Cat# 9644; RRID:AB_2097841 |
eIF4B | Cell Signaling Technology | Cat# 3591; RRID:AB_2097522 |
phospho-T37 4E-BP1 | Cell Signaling Technology | Cat# 2855; RRID:AB_560835 |
phospho-T70 4E-BP1 | Cell Signaling Technology | Cat# 9455; RRID:AB_330949 |
non-phospho-T46 4E-BP1 | Cell Signaling Technology | Cat# 4923; RRID:AB_659944 |
phospho-S65 4E-BP1 | Cell Signaling Technology | Cat# 9451; RRID:AB_330947 |
mTOR | Cell Signaling Technology | Cat# 2983; RRID:AB_2105622 #4517; RRID:AB_1904056 |
p70S6K1 | Cell Signaling Technology | Cat# 9234; RRID:AB_2269803 |
Ero-1Lα | Cell Signaling Technology | Cat# 3264; RRID:AB_823684 |
PDI | Cell Signaling Technology | Cat# 3501; RRID:AB_2156433 |
DCP1B | Cell Signaling Technology | Cat# 13233; RRID:AB_2798157 |
DDX6 | Cell Signaling Technology | Cat# 9407; RRID:AB_10556959 |
Tubulin | Cell Signaling Technology | Cat# 2128; RRID:AB_823664 |
CalR | Cell Signaling Technology | Cat# 2891; RRID:AB_2275208 |
PKCζ | Cell Signaling Technology | Cat# 9368; RRID:AB_10693777 |
PKC δ | Cell Signaling Technology | Cat# 2058; RRID:AB_10694655 |
BiP | Cell Signaling Technology | Cat# 3177; RRID:AB_2119845 |
LC3B | Cell Signaling Technology | Cat# 3868; RRID:AB_2137707 |
EEA1 | Cell Signaling Technology | Cat# 3288; RRID:AB_2096811 |
AIF | Cell Signaling Technology | Cat# 5318; RRID:AB_10634755 |
Rab5 | Cell Signaling Technology | Cat# 3547; RRID:AB_2300649 |
RCAS1 | Cell Signaling Technology | Cat# 12290; RRID:AB_2736985 |
Calnexin (CNX) | Cell Signaling Technology | Cat# 2679; RRID:AB_2228381 |
PTBP1/2 | Cell Signaling Technology | Cat# 8776 The product is discontinued |
hnRNPK | Cell Signaling Technology | Cat# 9081; RRID:AB_11178946 |
FUBP/ KHSRP (E2E2U) | Cell Signaling Technology | Cat# 13398; RRID:AB_2798208 |
IRE1α | Cell Signaling Technology | Cat# 3294; RRID:AB_823545 |
rpS6 | Cell Signaling Technology | Cat# 2217; RRID:AB_331355 |
rpL7a | Cell Signaling Technology | Cat# 2415; RRID:AB_2182059 |
SUN2 | Abcam | Cat# ab124916; RRID:AB_10972497 |
Nesprin 1/Syne-1 | Abcam | Cat# ab5250; RRID:AB_2200724 |
Exportin 5 | Abcam | Cat# ab57491; RRID:AB_941499 |
SUN2 | Sigma-Aldrich | Cat# MABT880 |
GRP78/HSPA5/BiP | Novus Biol | Cat# 06274; RRID:AB_1555284 |
eIF4E3 | Proteintech | Cat# 17282-1-AP; RRID:AB_2262162 |
Bacterial and virus strains | ||
ShRNA_eIF4E3 lentiviral particles | Santa Cruz Biotechnology | Cat# sc-78455-V |
Mammalian vector pCMV6-AC-GFP (Human) | Origene | Cat# PS100010 |
Chemicals, peptides, and recombinant proteins | ||
pp242 | Tocris Bioscience | Cat# 4257/5 |
Kinesin inhibitor K858 | Tocris Bioscience | Cat# 4960/1 |
Dynarrestin | Tocris Bioscience | Cat# 6526/5 |
Torin 1 | Tocris Bioscience | Cat# 4247/10 |
PF4708671 | Tocris Bioscience | Cat# 4032/5 |
Bio | Tocris Bioscience | Cat# 3194/10 |
Rotenone | Tocris Bioscience | Cat# 3616/50 |
MG132 | Tocris Bioscience | Cat# 1748/5 |
Geldanamycin | Tocris Bioscience | Cat# 1368/1 |
T4 Polynucleotide Kinase | New England Biolabs | Cat# M0201S |
T4 PK Reaction Buffer | New England Biolabs | Cat# B0201S |
DMEM | Thermo Fisher Scientific | Cat# A4192101 |
RPMI 1640 | Thermo Fisher Scientific | Cat# 61870143 |
FBS | Thermo Fisher Scientific | Cat# A4766 |
Penicillin/Streptomycin | Sigma | Cat# P4333 |
HEPES solution | Sigma | Cat# H0887 |
NaCl | Sigma | Cat# S5150 |
Glycerol | VWR | Cat# 24388-295 |
NP-40/IPEGAL | EMD Millipore | Cat# 492016 |
Triton X-100 | Sigma | Cat# T8787 |
Tris-HCl pH 7.5 1M | Thermo Fisher Scientific | Cat# 15567027 |
RNAsin RNase inhibitor | Promega | Cat# N251B |
Sodium deoxycholate | Ameresco | Cat# 0613 |
KCl | Thermo Fisher Scientific | Cat# AM9640G |
MgCl2 | Thermo Fisher Scientific | Cat# AM9530G |
LiCl | Invitrogen | Cat# L7026 |
DTT | OmniPur | Cat# 3860 |
Cycloheximide | Ameresco | Cat# 94271-5G |
RNase-free water | GE Healthcare | Cat# 0226 |
siRNA buffer | Dharmacon™ | Cat# B-002000-UB-100 |
Protease Inhibitors | Roche | Cat# 11393100 |
qScript cDNA Super-Mix | QuantaBio | Cat# 10142-784 |
Formaldehyde | Sigma | Cat# F9037 |
MNase | Sigma-Aldrich | Cat# N3755 |
Guanidine thiocyanate | Sigma | Cat# 50983 |
Trizol | Ambion | Cat# 10296-010 |
GlycoBlue | ThermoFisher Scientific | Cat# AM9515 |
Polynucleotide kinase | BioLabs | Cat# M0201S |
PBS | Gibco | Cat# 14200-075 |
Tween 20 | Sigma-Aldrich | Cat# H06J02 |
Puromycin | GIBCO | Cat# J67236.8EQ |
BSA | Sigma-Aldrich | Cat# A9647 |
Other | ||
Compound | Target | Concentration |
pp242 | mTOR | 500 nM |
K858 | Kinesin | 1–10 µM |
Dynarrestin | Dynein | 5–25 µM |
Torin 1 | mTOR | 250 nM |
PF4708671 | RSK | 5 µM |
Bio | GSK-3 | 1 µM |
Rotenone | Mitochondrial electron transport | 1 µM |
MG132 | 26S proteasome | 1 µM |
Geldanamycin | Hsp90 | 1 µM |
Critical commercial assays | ||
RNeasy Mini Kit | QIAGEN | Cat# B-002000-UB-100 |
Stellaris RNA FISH | Biosearch technologies | Cat# VSMF-20680-5 |
QIAseq miRNA Library Prep Kit | QIAGEN | cat #331502 |
MTT Cell Growth Assay Kit | Sigma-Aldrich | Cat# CT02 |
Deposited data | ||
mRNA and Ribo-seq data | NCBI GEO | GSE199535 |
The mass spectrometry proteomics data | ProteomeXchange Consortium | PRIDE: PXD031043 |
Experimental models: Cell lines | ||
Colorectal carcinoma HCT-15 | ATCC | CCL-225 |
Cervical adenocarcinoma HeLa | ATCC | CCL-2 |
Gastric adenocarcinoma AGS | ATCC | CRL-1739 |
Hepatocellular carcinoma Hep G2 | ATCC | HB-8065 |
Experimental models: Organisms/strains | ||
HCT-15, GFP- rpL7a expression | Origene | Cat# PS100010 |
HeLa, siRNA gene silencing, SUN2 | Dharmacon™ | Cat# 25777 |
HCT-15, Transduction of shRNA eIF4E3 lentiviral particles | Santa Cruz Biotechnology | Cat# sc-78455-V |
Oligonucleotides | ||
Human SUN2 siRNA | Dharmacon™ | Cat# 25777 |
ShRNA_eIF4E3 lentiviral particles | Santa Cruz Biotechnology | Cat# sc-78455-V |
Software and algorithms | ||
Proteome Discoverer 2.1 software | Thermo Scientific | https://www.thermofisher.com/us/en/home/industrial/mass-spectrometry/liquid-chromatography-mass-spectrometry-lc-ms/lc-ms-software/multi-omics-data-analysis/proteome-discoverer-software.html |
Mascot 2.4 | Matrix Science | https://www.matrixscience.com/ |
gpGrouper algorithm | Saltzman et al. (2018, MCP)85 | PMID: 30093420 |
Gene Set Enrichment Analysis (GSEA) | Subramanian et al. (2005, PNAS)86 | https://www.gsea-msigdb.org/gsea/index.jsp |
FastQC | Andrews et al. (2010)87 | https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ |
Trimmomatic (v0.38) | Bolger et al., 201488 | Bioinformatics 30, 2114-2120 |
Bowtie aligner (v1.3.0) | Langmead et al., 200989 | Genome Biology 10, R25. |
The STAR aligner (v2.7.6a) | Dobin et al., 201390 | Bioinformatics 29, 15-21. |
R package edgeR | Robinson et al., 201091 | Bioinformatics 26, 139-140 |
RUV normalization | Risso et al., 201492 | Nature biotechnology 32, 896-902 |
MSigDB | Liberzon et al., 201193 | Bioinformatics 27, 1739-1740 |
MATLAB | Mathworks | https://www.mathworks.com/products/matlab.html |
Image Studio Lite | LI-COR | https://www.licor.com/bio/image-studio-lite/ |
GraphPad Prism 9.4.1 | GraphPad | https://www.graphpad.com/scientific-software/prism/ |
Adobe Illustrator CC 2022 | Adobe | https://www.adobe.com/products/illustrator.html |
NCBI GEO RNA-seq data | GEO accession GSE199535 | https://www.ncbi.nlm.nih.gov/gds |
ProteomeXchange Proteomic data | Dataset Identifier PXD 031043 |
http://proteomecentral.proteomexchange.org/cgi/GetDataset |