REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Rabbit polyclonal anti LARP6	Atlas Antibodies	Cat#HPA049029; RRID: AB_2680604
Mouse monoclonal anti vimentin	Abcam	Cat#ab8978; RRID: AB_306907
Rabbit polyclonal anti GFP	Abcam	Cat#ab290; RRID: AB_303395
GFP-Trap®	ChromoTek	Cat#gtma-20; RRID: AB_2631358
Mouse monoclonal anti E-cadherin	Cell signalling	Cat#3195; RRID: AB_2291471
Mouse monoclonal anti TCF8/ZEB	Cell signalling	Cat#3396; RRID: AB_1904164
Phospho-p70 S6 Kinase (Thr389)	Cell signalling	Cat#9206; RRID: AB_2285392
p70 S6 Kinase	Cell signalling	Cat#2708; RRID: AB_390722
α-Tubulin (DM1A)	Cell signalling	Cat#3873; RRID: AB_1904178
α-Tubulin (11H10)	Cell signalling	Cat#2125; RRID: AB_2619646
α-Tubulin Monoclonal Antibody	Thermo Fisher	Cat#A11126; RRID: AB_221538
GAPDH	Novus Biologicals	Cat#NB300-221;RRID: AB_10077627
Alexa Fluor™ 488 Phalloidin	Thermo Fisher Scientific	Cat#A12379
Rabbit IgG HRP linked	GE Healthcare	Cat#NA934; RRID: AB_772206
Mouse IgG HRP linked	GE Healthcare	Cat#NA931; RRID: AB_772210
Bacterial and Virus Strains
DH5α for cloning	Thermo Fisher Scientific	Cat#18265017
Chemicals, Peptides, and Recombinant Proteins
NuPAGE LDS Sample Buffer	Thermo Fisher Scientific	Cat#NP0008
Pierce ECL Plus Western Blotting Substrate	Thermo Fisher Scientific	Cat#N32132
Click-iT™ Plus OPP Alexa Fluor™ 488 Protein Synthesis Assay	Thermo Fisher Scientific	Cat#C10456
Horse Serum	Invitrogen	Cat#16050-122
Human EGF	Prepotech	Cat#AF-100-15-1000
Hydrocortisone	Sigma-Aldrich	Cat#H0888; CAS: 50-23-7
Cholera Toxin from Vibrio cholerae	Sigma-Aldrich	Cat#C8052; CAS: 9012-63-9
Insulin from bovine pancreas	Sigma-Aldrich	Cat#I1882; CAS: 11070-73-8
Crystal violet	Sigma-Aldrich	Cat#C6158; CAS: 548-62-9
DTT	VWR Chemicals	Cat#M109; CAS: 3483-12-3
Iodoacetamide	VWR Chemicals	Cat#786-228; CAS: 144-48-9
Blasticidin S HCl	Life Technologies	Cat#R21001
TGF-β1 human	Sigma-Aldrich	Cat#H8541
Emetine	Sigma-Aldrich	Cat#E2375; CAS: 7083-71-8
Harringtonine	Sigma-Aldrich	Cat#SML1091; CAS: 26833-87-4
Puromycin	Sigma-Aldrich	Cat#P9620; CAS: 58-58-2
ANTI-RNase (15-30 U/μL)	Life Technologies Ltd Invitrogen Division	Cat#AM2692
RNase A, DNase and protease-free (10 mg/mL)	Life Technologies Ltd	Cat#EN0531
AZD8055	Selleckchem	S1555; CAS: 1009298-09-2
Torin	Selleckchem	S2827; CAS: 1009298-09-2
Everolimus	Selleckchem	S1120; CAS: 159351-69-6
Hoechst 33342	Thermo Fisher Scientific	Cat#H3570
HCS NuclearMask™ Blue Stain	Thermo Fisher Scientific	Cat#H10325
C9 (Stefanovic et al., 2019)	ChemBridge	DIVERSet-CL chemical library
SignalStain® Antibody Diluent	Cell Signaling Technologies	Cat#8112S
SignalStain® DAB Substrate Kit	Cell Signaling Technologies	Cat#8059P
SignalStain® Boost IHC Detection Reagent (HRP, Rabbit)	Cell Signaling Technologies	Cat#8114P
Antigen Unmasking Solution, Citric Acid Based	Vector Laboratories	Cat#H-3300
DPX new	Merck	Cat#100579
RNAscope® Fluorescent Multiplex Reagent Kit	Advanced Cell Diagnostics Srl	Cat#320850
Hs-RPL34 targeting 24-702 of NM_000995.4	Advanced Cell Diagnostics Srl	Cat#504031
Hs-RPS7 targeting 2-521 of NM_001011.3	Advanced Cell Diagnostics Srl	Cat#504211
Hs-RPLP2	Advanced Cell Diagnostics Srl	Cat#511391
Hs-RPL22	Advanced Cell Diagnostics Srl	Cat#435271
Hs-RPS21	Advanced Cell Diagnostics Srl	Cat#511381
Hs-ITGB4	Advanced Cell Diagnostics Srl	Cat#300031
Edit-R CRISPRa crRNA Non-targeting control, 5nmol	Dharmacon	Cat#U-009500-01-05
Edit-R CRISPR-Cas9 Synthetic tracrRNA, 5 nmol	Dharmacon	Cat#U-002005-05
Edit-R Modified Synthetic crRNA, desalted/deprotected, 2 nmol: LARP6; chr15:70832948(+) - custom design	Dharmacon	Cat#crRNA-409669
DharmaFECT Duo	Dharmacon	Cat#T-2010-01
CellTracker™ Green CMFDA Dye	Thermo Fisher Scientific	Cat#C7025
CellTracker™ Orange CMTMR Dye	Thermo Fisher Scientific	Cat#C2927
Collagen I	Advanced BioMatrix	Cat#5005
Lipofectamine 2000 Transfection Reagent-1.5 mL	Life Technologies	Cat#11668019
Lipofectamine™ RNAiMAX Transfection Reagent	Thermo Fisher Scientific	Cat#13778150
Opti-MEM I Reduced Serum Medium-100 mL	Thermo Fisher Scientific	Cat#31985062
Gibco™ DMEM w/High Glucose and w/o Glutamine, Lysine and Arginine	Fisher Scientific	Cat#12817552
Critical Commercial Assays
TMTsixplex™ Isobaric Label Reagent Set	Thermo Fisher Scientific	Cat#90061
TMT10plex™ Isobaric Label Reagent Set	Thermo Fisher Scientific	Cat#90110
Subcellular Protein Fractionation Kit	Thermo Fisher Scientific	Cat#78840
Pierce High pH Reversed-Phase Peptide Fractionation Kit	Life Technologies	Cat#84868
QuantSeq mRNA 3’ end sequencing kit	Lexogen	Cat#SKU: 015.24
CellTiter-Glo® Luminescent Cell Viability Assay	Promega	Cat#G7571
RNeasy Mini Kit	QIAGEN	Cat#74104
MTT Cell Viability Assay	Thermo Fisher Scientific	Cat#M6494
Qubit™ RNA HS Assay Kit	Thermo Fisher Scientific	Cat#Q32852
Pierce™ BCA Protein Assay Kit	Thermo Fisher Scientific	Cat#23225
High Sens. RNA ScreenTape Sample Buffer	Agilent Technologies	Cat#5067-5580
High Sensitivity RNA ScreenTape	Agilent Technologies	Cat#5067-5579
Brilliant II SYBR® Green QRT-PCR	Agilent Technologies	Cat#600825
MycoAlert™ PLUS Mycoplasma Detection Kit	Lonza	Cat#LT07-705
Deposited Data
TMT quantitative proteomics analysis of (a) protrusions and cells bodies of MDA-MB231 cells collected after 1, 2, 4, & 8 hrs post protrusion induction, and (b) protrusions and cells bodies of a panel of five normal and malignant human cell-lines.	This paper	PXD021239 accessible via PRIDE partner repository (http://www.ebi.ac.uk/pride/archive/)
TMT quantitative proteomics analysis of MDA-MB231 cells grown on either closed or open pore 3 μm transwells for either 2 or 24 hrs.	This paper	PXD021206 accessible via PRIDE partner repository (http://www.ebi.ac.uk/pride/archive/)
Pulsed SILAC quantification of translation rates between MDA-MB231 cells grown on either closed or open pore 3 μm transwells for 1, 2, 4 or 8 hrs.	This paper	PXD021203 accessible via PRIDE partner repository (http://www.ebi.ac.uk/pride/archive/)
Pulsed SILAC coupled with iBAQ quantification of protein abundances within different subcellular locations in MDA-MB231 cells, following protrusion induction.	This paper	PXD021205 accessible via PRIDE partner repository (http://www.ebi.ac.uk/pride/archive/)
TMT quantitative proteomics analysis of non-targeted control siRNA (NT) or LARP6 siRNA transfected MDA-MB231 cells, grown overnight on either closed (no protrusions) or open pore (with protrusions) 3 μm transwells.	This paper	PXD021180 accessible via PRIDE partner repository (http://www.ebi.ac.uk/pride/archive/)
SILAC proteomics analysis of non-targeted (NT) vs LARP6 siRNA treated MDA-MB231 cells.	This paper	PXD021204 accessible via PRIDE partner repository (http://www.ebi.ac.uk/pride/archive/)
3’ mRNA-seq (QUANTSEQ FWD) sequencing of protrusion and cell body fractions of BJ, PC-3M, RPE-1, U-87 and WM-266.4 cells.	This paper	E-MTAB-8470 accessible via ArrayExpress repository (http://www.ebi.ac.uk/arrayexpress)
3’ mRNA-seq (QUANTSEQ FWD) sequencing of protrusion and cell body fractions of non-transfected control siRNA or LARP6 siRNA transfected MDA-MB231 cells.	This paper	E-MTAB-9520 accessible via ArrayExpress repository (http://www.ebi.ac.uk/arrayexpress)
GFP-LARP6 and GFP control sequencing of iCLIP libraries generated from stably expressing MDA-MB231 cells.	This paper	E-MTAB-9636 Accessible via ArrayExpress repository (http://www.ebi.ac.uk/arrayexpress)
Experimental Models: Cell Lines
MDA-MB231	ATCC	ATCC number 92020424
RPE-1	N/A	Dr Sarah McClelland (Barts Cancer Institute)
BJ	N/A	Dr Sarah McClelland (Barts Cancer Institute)
U-87	N/A	Dr Paul Huang (Institute of Cancer Research)
WM-266.4	N/A	Prof. Chris Marshall (Institute of Cancer Research)
PC-3M	N/A	Dr Prabhakar Rajan (Barts Cancer Institute)
MCF10AT	N/A	Dr Susana Godinho (Barts Cancer Institute)
Oligonucleotides
SIRNA UNIV NEGATIVE CONTROL	Sigma-Aldrich	Cat#SIC001
ON-TARGETplus Non-targeting Pool	Dharmacon	Cat#D-001810-10-05
LARP6 siRNA-1 GGAUUCAUGGCCAUGAGA	Sigma MISSION	Cat#Hs02_00351818
LARP6 siRNA-2 GCAAGAUGCUCCUGGUCUA	Sigma MISSION	Cat#Hs01_00153597
LARP6 siRNA-3 CUGUGUAUAAAUACCUUCU	Sigma MISSION	Cat#Hs01_00153598
LARP1 siRNA CUGACUAUGAGAUUGAUGA	Sigma MISSION	Cat#Hs01_00168468
LARP1B siRNA GAGAAUGAUACACGAAGU; AGACCUGGAUCCCGGAACA; UCAAGUAAUCAACGUAAGA; GGUGGUAAUAUCCGAGGUU	Dharmacon	SMARTpool: ON-TARGETplus L-013350-02-0005
LARP7 siRNA AGGAAACAGUCCGGGAUA; GUGCUAUCAAAGAGCGAAU; GCAAAGACUCAACAAGCGA; CUUAAUCAGCCUCGGGAAA	Dharmacon	SMARTpool: ON-TARGETplus L-020996-01-0005
SSB siRNA GGUCGUAGAUUUAAAGGAA; GGUUAGAAGAUAAAGGUCA; GAGACCAGUAGUUUAGUAA; GGGAAGUACUAGAAGGAGA	Dharmacon	SMARTpool: ON-TARGETplus L-006877-01-0005
h47S rRNA fw 5’ TTCGTTCGCTCGCTCGTT 3’	Sigma	N/A
h47S rRNA rv 5’ CAACGACACGCCCTTCTTTC 3’	Sigma	N/A
Hs_LARP6_1_SG QuantiTect Primer Assay	QIAGEN	Cat#QT00221445
Hs_GAPDH_1_SG QuantiTect Primer Assay	QIAGEN	Cat#QT00079247
Recombinant DNA
TetO-WT-L32TOP-β-Globin-12xMS2 (WT 5’TOP reporter)	Gift from A. Gentilella	N/A
TetO-MUT-L32TOP-β-Globin-12xMS2 (MUT 5’TOP reporter)	Gift from A. Gentilella	N/A
MCP-EGFP expression plasmid	Gift from C. Gallego	N/A
VSV lentiviral packaging vectors	Gift from C. Gallego	N/A
deltaR lentiviral packaging vectors	Gift from C. Gallego	N/A
rtTA-N144 plasmid (Richner et al., 2015)	Gift from A. Yoo	Cat#66810; RRID: Addgene_66810
pTK-Twist lentiviral inducible expression plasmid (Guo et al., 2012)	Gift from B. Weinberg	Cat#36977; RRID: Addgene_3697
pTK-Snail lentiviral inducible expression plasmids (Guo et al., 2012)	Gift from B. Weinberg	Cat#36976; RRID: Addgene_3697
pcDNA™6.2/N-EmGFP-DEST Vector	Thermo Fisher Scientific	Cat#V35620
Software and Algorithms
MaxQuant	N/A	https://www.biochem.mpg.de/5111795/maxquant
Perseus	N/A	https://www.biochem.mpg.de/5111810/perseus
BlueBee	N/A	https://www.bluebee.com/
Galaxy	N/A	https://usegalaxy.org/
GradPad PRISM v7	N/A	https://www.graphpad.com/scientific-software/prism/
iMAPS webserver	N/A	https://imaps.genialis.com/iclip
ImageJ	N/A	https://imagej.net/
Other
75 mm Transwell with 3.0 μm pore polycarbonate membrane insert	Corning	Cat#3420
24 mm Transwell with 3.0 μm pore polycarbonate membrane insert	Corning	Cat#3414
6.5 mm Transwell with 3.0 μm pore polycarbonate membrane insert	Corning	Cat#3415
Vivacon 500, 30,000 MWCO Hydrosart	Sartorius	Cat#VN01H22
iBiDi μ-Slide 18 Well flat, ibiTreat	Thistle Scientific	Cat#81826
iBiDi μ-Plate 96 Well Black	Thistle Scientific	Cat#89626
Falcon™ Chambered Cell Culture Slides	Thermo Fisher Scientific	Cat#354118