REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
GAPDH	Cell signaling	Cat#2118; RRID:AB_561053
β-actin	Cell signaling	Cat#58169; RRID:AB_2750839
HA	Cell signaling	Cat#3724; RRID:AB_1549585
HA	Cell signaling	Cat#2367; RRID:AB_10691311
V5	Cell signaling	Cat#13202; RRID:AB_2687461
Cleaved-PARP	Cell signaling	Cat#5625; RRID:AB_10699459
GFP	Cell signaling	Cat#2555; RRID:AB_10692764
FLAG	Cell signaling	Cat#14793; RRID:AB_2572291
ERG	Cell signaling	Cat#97249; RRID:AB_2721841
FLI	Abcam	Cat#15289; RRID:AB_301825
NKX2-2	Abcam	Cat#187375; RRID:AB_2889166
FLAG-M2	Sigma	Cat#F3165; RRID:AB_259529
TRIM8	Santa Cruz Biotechnology	Cat#sc-398878; RRID:AB_2889167
EWS	Santa Cruz Biotechnology	Cat#sc-28327; RRID:AB_675526
FLAG	Novus Biologicals	Cat#NBP1-06712; AB_1625981
Ubiquitin	LifeSensors	Cat#VU191; RRID:AB_2716558
Bacterial and virus strains
One Shot™ Stbl3 E.coli	ThermoFisher Scientific	Cat#C737303
Biological samples
EW_PDX001	Garcia-Dominguez et al.2018 (PMID: 30140378)	N/A
Chemicals, peptides, and recombinant proteins
Lipofectamine RNAiMAX	ThermoFisher Scientific	Cat#13778030
dTAGV-1	Nabet et al., 2020 (PMID: 32948771)	N/A
Dynabeads™ Protein G	Invitrogen	Cat#10004D
Dynabeads™ Protein A	Invitrogen	Cat#10001D
BD Pharmingen™ Transcription-Factor Buffer Set	FisherScientific	Cat#BDB562574
FuGENE® HD Transfection Reagent	Promega	Cat#E2311
UM402M: TUBE 2 (Magnetic Beads)	LifeSensors	Cat#UM402M
Janelia Fluor® 646 HaloTag® Ligands	Promega	Cat#GA1120
Duolink® In Situ PLA® Probe Anti-Mouse MINUS Affinity purified Donkey anti-Mouse IgG (H+L)	Sigma	Cat#DUO92004
Duolink® In Situ PLA® Probe Anti-Rabbit PLUS Affinity purified Donkey anti-Rabbit IgG (H+L)	Sigma	Cat#DUO92002
Critical commercial assays
CellTiter-Glo® 2.0 Cell Viability Assay	Promega	Cat#G9241
Duolink™ flowPLA Detection Kit - Green	Sigma	Cat#DUO94002
Click-iT™ EdU Pacific Blue™ Flow Cytometry Assay Kit	ThermoFisher Scientific	Cat#C10418
Deposited data
Raw and analyzed data	This study	GEO: GSE150244
Flow cytometry-based CRISPR screen	This study	Table S1
Proteomics experiment	This study	Table S2
DepMap 20Q1 genome-scale CRISPR-Cas9 screening and cell line characterization data	Broad Institute DepMap	https://depmap.org; https://figshare.com/articles/DepMap_20Q1_Public/11791698/3
Experimental models: cell lines
EWS502 N-dTAG E/F	Nabet et al., 2020(PMID: 32948771)	N/A
TC32 N-dTAG TRIM8	This study	N/A
TC32 C-dTAG TRIM8	This study	N/A
TC71 N-dTAG TRIM8	This study	N/A
TC71 C-dTAG TRIM8	This study	N/A
293T EWS/FLI-GFP-IRES-mCherry-1	This study	N/A
Experimental models: organisms/strains
NU/NU Nude Mouse - Crl:NU-Foxn1nu	Charles River	Cat#088; RRID:IMSR_CRL:088
NGS Mouse - NOD-scid IL2Rgammanull	The Jackson Laboratory	Cat#005557; RRID:IMSR_JAX:005557
Oligonucleotides
sgNT: 5’ GTAGCGAACGTGTCCGGCGT3’	This study	N/A
sgTRIM8-1: 5’ CTACCGCCTCTACCACTGCG 3’	This study	N/A
sgTRIM8-2: 5’ GCACGTGGAGAAGCCGCCGG 3’	This study	N/A
sgTRIM8-3: 5’ GCGCAGGCAGACCTTCTGCG 3’	This study	N/A
ON-TARGETplus Non-targeting Control siRNA	Dharmacon	Cat#001810
SMARTpool TRIM8 siRNA	Dharmacon	Cat#81603
Recombinant DNA
Artichoke-EWS/FLI (EWS/FLI-GFP-IRES-mCherry)	This study	N/A
pLEX305-N-dTAG-EWS/FLI	Nabet et al., 2020 (PMID: 32948771)	N/A
pLEX305-N-dTAG-TRIM8	This study	N/A
pLEX305-C-dTAG-TRIM8	This study	N/A
pLX307-TRIM8-V5	This study	N/A
pLX307-TRIM8ΔRING-V5	This study	N/A
pLX307-TRIM8ΔB-box1-V5	This study	N/A
pLX307-TRIM8ΔB-box2-V5	This study	N/A
pLX307-TRIM8ΔCC-V5	This study	N/A
pLX307-TRIM8ΔRFP-like-V5	This study	N/A
pLX307-TRIM8-EGFP	This study	N/A
pLX307-EWS/FLI-3XFLAG	This study	N/A
pLX307-N-EWS-3XFLAG	This study	N/A
pLX307-C-FLI-3XFLAG	This study	N/A
pLX307-EWS/ERG-3XFLAG	This study	N/A
pLX307-ERG-3XFLAG	This study	N/A
pLX307-N1-FLI-3XFLAG	This study	N/A
pLX307-N2-FLI-3XFLAG	This study	N/A
pLX307-EWS/FLIΔC1-3XFLAG	This study	N/A
pLX307-EWS/FLIΔC2-3XFLAG	This study	N/A
pLX307-EWS/FLIΔC3-3XFLAG	This study	N/A
pLX307-EWS/FLIΔC4-3XFLAG	This study	N/A
pLX307-EWS/FLIΔC5-3XFLAG	This study	N/A
pLX307-EWS-C1-3XFLAG	This study	N/A
pLX307-EWS-C5-3XFLAG	This study	N/A
pLX307-EWS/FLI-K144R-3XFLAG	This study	N/A
pLX307-EWS/FLI-K334R-3XFLAG	This study	N/A
pLX307-EWS/FLI-K217R-3XFLAG	This study	N/A
pLX307-EWS/FLI-K240R-3XFLAG	This study	N/A
pLX307-EWS/FLI-K217/240R-3XFLAG	This study	N/A
pLX307-EWS/FLIΔC3-K144R-3XFLAG	This study	N/A
pLX307-HA-Ubiquitin	This study	N/A
pLIV-V5-EWS-FLI1	Boulay et al., 2017 (PMID: 28844694)	N/A
pINDUCER20-EWS/FLI-HA	This study	N/A
Software and algorithms
limma v3.42.2 R package	Ritchie et al., 2015 (PMID: 25605792)	https://bioconductor.org/packages/release/bioc/html/limma.html
FASTQC v0.11.9 software	N/A	www.bioinformatics.babraham.ac.uk/projects/fastqc/
multiqc v1.6 tool	Ewels et al., 2016 (PMID: 27312411)	https://multiqc.info
STAR v2.7.2b	Dobin et al., 2013 (PMID: 23104886)	https://github.com/alexdobin/STAR
ercc dashboard R package available from Bioconductor v3.10	Munro et al., 2014 (PMID: 25254650)	https://bioconductor.org/packages/release/bioc/html/erccdashboard.html
SARTools v1.7.3	Varet et al., 2016 (PMID: 27280887)	https://github.com/PF2-pasteur-fr/SARTools
featureCounts v1.6.3 method implemented in the Subread v2.0.0 R package	Liao et al., 2014 (PMID: 24227677)	http://subread.sourceforge.net
DESeq2 v1.24.0	Love et al., 2014 (PMID: 25516281)	https://bioconductor.org/packages/release/bioc/html/DESeq2.html
enricher function in the clusterProfiler v 3.14.3 R package	Yu et al., 2012 (PMID: 22455463)	https://www.bioconductor.org/packages/release/bioc/html/clusterProfiler.html
Proteome Discoverer 2.2	ThermoFisher	RRID:SCR_014477
Other
Code used to analyze data and generate figures	This study	https://github.com/ndharia-broad/TRIM8_Ewing DOI: https://doi.org/10.6084/m9.figshare.14888382.v1