Antibodies |
chicken anti-GFP |
Abcam |
ab13970; RRID:AB_300798 |
mouse anti-BrdU (MoBU-1) |
Invitrogen |
B35128; RRID:AB_2536432 |
rabbit anti-Keratin 10 (Poly19054) |
BioLegend |
905403; RRID:AB_2749902 |
mouse anti-Puromycin (12D10) |
Millipore |
MABE343; RRID:AB_2566826 |
guinea pig anti-Keratin 5 |
Origene |
BP5006 |
CD49f/α6-Integrin-PerCP (GoH3) |
BioLegend |
313617 RRID:AB_1575054 |
CD326/EpCAM-APC (G8.8) |
BD Biosciences |
563478; RRID:AB_2738234 |
mouse anti-EIF2B5 (B-7) |
Santa Cruz Biotechnology |
Sc-55558; RRID:AB_831253 |
mouse anti-β-Actin (2D4H5) |
Proteintech |
66009; RRID:AB_2687938 |
mouse anti-EIF2α (L57A5) |
Cell Signaling |
2103S; RRID:AB_836874 |
rabbit anti-phospho-EIF2α Ser51 (119A11) |
Cell Signaling |
3597S; RRID:AB_390740 |
rabbit anti-FBXO32 (EPR9148(2)) |
Abcam |
ab168372 |
mouse anti-V5 (V5-10) |
Sigma-Aldrich |
V8012; RRID:AB_261888 |
Bacterial and Virus Strains |
TRC1.0 Lentiviral Mouse Genome shRNA Library |
Sigma-Aldrich |
Discontinued, now MISSION shRNA Library |
Biological Samples |
|
|
|
|
|
Chemicals, Peptides, and Recombinant Proteins |
DMEM |
Gibco |
11995-065 |
UltraCULTURE media |
Lonza |
12-725F |
E media |
Nowak and Fuchs, 2009 |
N/A |
Fetal bovine serum |
Gibco |
16000-044 |
Polybrene |
Sigma |
107689-10G |
Trypsin-EDTA |
GIBCO |
25200-056 |
Chelex 100 |
Bio-Rad |
1422822 |
M.O.M. Blocking Reagent |
Vector |
MKB-22113 |
ProLong Gold Antifade Mountant |
ThermoFisher |
P36930 |
Antigen Unmasking Solution, Citric Acid Based |
Vector |
H-3300-250 |
RIPA Lysis Buffer System |
Santa Cruz Biotech |
Sc-24948 |
ISRIB |
Sigma-Aldrich |
SML0843 |
SuperSignal Wester Pico PLUS Chemiluminescent Substrate |
Thermo Fisher |
34577 |
iScript Reverse Transcription Supermix |
Bio-Rad |
1708840 |
SYBR Green PCR Master Mix |
Thermo Fisher |
4309155 |
Lipofectamine 3000 |
Thermo Fisher |
L3000001 |
BrdU |
Invitrogen |
B23151 |
EdU |
Invitrogen |
A10044 |
Puromycin |
Thermo Fisher |
A11138-03 |
Cyclohexamide |
Sigma-Aldrich |
C7698 |
Bortezomib |
Sigma-Aldrich |
504314 |
Dispase |
Corning |
354235 |
Mitomycin C |
Sigma-Aldrich |
M4287 |
Critical Commercial Assays |
Click-iT EdU Alexa Fluor 555 Imaging Kit |
Invitrogen |
C10338 |
20S Proteasome Activity Assay |
Millipore |
APT280 |
PE Active Caspase-3 Apoptosis Kit |
BD Biosciences |
550914 |
DNeasy Blood & Tissue Kit |
Qiagen |
69504 |
RNeasy Plus Mini Kit |
Qiagen |
74134 |
Q5 Site-Directed Mutagenesis Kit |
NEB |
E0554S |
Pierce BCA Protein Assay Kit |
Thermo Fisher |
23225 |
Dual-Luciferase Reporter Assay System |
Promega |
E1910 |
TruSeq Ribo Profile (Mammalian) Library Prep Kit |
Illumina |
RPHMR12126 |
Ribo-Zero rRNA Removal Kit (Human/Mouse/Rat) |
Illumina |
MRZH11124 |
Deposited Data |
Raw and analyzed data |
This paper |
GEO GSE 126660 |
Mouse reference genome NCBI build 38 (mm10) |
Genome Reference Consortium |
https://www.ncbi.nlm.nih.gov/assembly/GCF_000001635.20/ |
Experimental Models: Cell Lines |
Mouse: WT primary keratinocytes |
This paper |
N/A |
Mouse: HrasG12V primary keratinocytes |
This paper |
N/A |
Mouse: NIH/3T3 cells |
ATCC |
CRL-1658; RRID:CVCL_0594 |
Human: 293TN cells |
System Biosciences |
LV900A-1 |
Experimental Models: Organisms/Strains |
Mouse: Rosa26YFPFYP
|
The Jackson Laboratory |
006148; RRID:IMSR_JAX:006148 |
Mouse: Rosa26mT/mG
|
The Jackson Laboratory |
007676; RRID:IMSR_JAX:007676 |
Mouse: FR-HrasG12V
|
Donated by James A. Fagin |
N/A |
Oligonucleotides |
See Table S6 for shRNA and PCR primer sequences |
|
|
Recombinant DNA |
Plasmid: pLKO.1 |
Addgene |
10878; RRID:Addgene_10878 |
Plasmid: pLKO.1-Cre |
Addgene |
25997; RRID:Addgene_25997 |
Plasmid: pLX-Cre-EF1α ORF vector |
Ying et al., 2018 |
N/A |
Plasmid: pLX-shRNA-Cre-EF1α ORF vector |
This paper |
N/A |
Plasmid: pMD2.G |
Addgene |
12259; RRID:Addgene_12259 |
Plasmid psPAX2 |
Addgene |
12260; RRID:Addgene_12260 |
Plasmid: CMV-Luc2CP/ARE |
Addgene |
62857; RRID:Addgene_62857 |
Software and Algorithms |
FASTX-Toolkit |
Hannon Lab |
http://hannonlab.cshl.edu/fastx_toolkit/ |
BWA |
Li et al., 2009 |
http://bio-bwa.sourceforge.net/ |
Samtools |
Li et al., 2009 |
http://samtools.sourceforge.net/ |
DESeq2 |
Love et al., 2014 |
https://bioconductor.org/packages/release/bioc/html/DESeq2.html |
Bowtie |
Langmead et al., 2009 |
https://bioconductor.org/packages/release/bioc/html/Rbowtie.html |
TopHat2 |
Kim et al., 2013 |
https://ccb.jhu.edu/software/tophat/index.shtml |
HTSeq |
Anders et al., 2015 |
https://htseq.readthedocs.io/en/release_0.10.0/ |
Xtail |
Xiao et al., 2016 |
https://github.com/xryanglab/xtail |
edgeR |
Robinson et al., 2010 |
https://bioconductor.org/packages/release/bioc/html/edgeR.html |
riboseqR |
Chung et al., 2015 |
http://bioconductor.org/packages/release/bioc/html/riboSeqR.html |
Ingenuity Pathway Analysis |
Qiagen |
https://www.qiagenbioinformatics.com/products/ingenuity-pathway-analysis/ |
Imaris |
Bitplane |
https://imaris.oxinst.com/ |
ImageJ |
Abramoff et al., 2004 |
https://imagej.nih.gov/ij/ |
Prism 7 |
GraphPad |
https://www.graphpad.com/ |
MEME |
Bailey et al., 2009 |
http://meme-suite.org/tools/meme |
FIMO |
Grant et al., 2011 |
http://meme-suite.org/tools/fimo |
ZEN |
Zeiss |
https://www.zeiss.com/microscopy/us/products/microscope-software/zen-lite.html |