Antibodies |
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Rabbit anti-AFF1 Antibody, Affinity Purified |
Bethyl Laboratories |
Bethyl Cat# A302-344A, RRID:AB_1850255 |
Rabbit anti-ELL2 Antibody, Affinity Purified |
Bethyl Laboratories |
Bethyl Cat# A302-505A, RRID:AB_1966087 |
Pol II Rpb1 NTD (D8L4Y) Rabbit mAb |
Cell Signaling Technology |
Cell Signaling Technology Cat# 14958, RRID:AB_2687876 |
BRD2 (D89B4) Rabbit mAb |
Cell Signaling Technology |
Cell Signaling Technology Cat# 5848, RRID: AB_10835146 |
BRD4 (E2A7X) Rabbit mAb |
Cell Signaling Technology |
Cell Signaling Technology Cat# 13440, RRID: AB_2687578 |
MED26 (D4B1X) Rabbit mAb |
Cell Cell Signaling Technology |
Cell Signaling Technology Cat# 14950 |
c-Myc (D3N8F) Rabbit mAb |
Cell Signaling Technology |
Cell Signaling Technology Cat# 13987, RRID:AB_2631168 |
Tubulin beta antibody |
DSHB |
DSHB Cat# E7, RRID:AB_528499 |
AFF4 Antibody |
Proteintech |
Proteintech Group Cat# 14662–1-AP, RRID:AB_2242609 |
Cyclin T1 Antibody (H-245) |
Santa Cruz |
Santa Cruz Biotechnology Cat# sc-10750, RRID:AB_2073888 |
HSP 90α/β Antibody |
Santa Cruz |
Santa Cruz Biotechnology Cat# sc-7947, RRID:AB_2121235 |
CDK9 Antibody (C-20) |
Santa Cruz |
Santa Cruz Biotechnology Cat# sc-484, RRID:AB_2275986 |
Anti-RNA polymerase II subunit B1 (phospho CTD Ser2) Antibody, clone 3E10 |
Active Motif |
Active Motif Cat# 61083, RRID:AB_2687450 |
FLAG-synthetic antibody |
Sigma-Aldrich |
Sigma-Aldrich Cat# F3165, RRID:AB_259529 |
ANTI-FLAG M2 Affinity Gel |
Sigma-Aldrich |
Sigma-Aldrich Cat# A2220, RRID:AB_10063035 |
Anti-SPT5 Antibody, clone 6F1 |
Millipore |
Millipore Cat# MABE1803 |
Rabbit anti-AFF1 serum |
(Lin et al., 2010) |
N/A |
Rabbit anti-ELL2 serum |
(Lin et al., 2010) |
N/A |
Rabbit anti-AFF4 serum |
(Lin et al., 2010) |
N/A |
Rabbit anti-Drosophila Rpb1 antibody |
(Lin et al., 2010) |
N/A |
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Bacterial and Virus Strains |
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Biological Samples |
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Chemicals, Peptides, and Recombinant Proteins |
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Flavopiridol |
Cayman |
Cat# 10009197 |
4-Thiouridine |
Sigma-Aldrich |
Cat# T4509 |
Screening compounds |
ChemDiv, ChemBridge and Enamine |
N/A |
EZ-Link™ HPDP-Biotin |
Thermo |
Cat# 21341 |
KL-1 |
This study |
N/A |
KL-2 |
This study |
N/A |
Biotin-AFF4 (32–67) peptides |
VCPBIO |
N/A |
Biotin-AFF4 mutant peptides |
VCPBIO |
N/A |
Glycogen |
Sigma-Aldrich |
Cat# 10901393001 |
GST-CCNT1 (1–300) |
This study |
N/A |
α-Amanitin |
Santa Cruz |
CAS 23109–05-9 |
Biotin-11-ATP |
Perkin Elmer |
Cat# NEL544001EA |
Biotin-11-CTP |
Perkin Elmer |
Cat# NEL542001EA |
Biotin-11-GTP |
Perkin Elmer |
Cat# NEL545001EA |
Biotin-11-UTP |
Perkin Elmer |
Cat# NEL543001EA |
RNA 5’ Pyrophosphohydrolase (RppH) |
NEB |
Cat# M0356S |
2% Agarose, PippinHT, 100–600 bp.10/pkg. |
Sage science |
Cat# HTC2010 |
BD Matrigel Matrix |
BD Biosciences |
Cat# 354234 |
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Critical Commercial Assays |
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TruSeq® Stranded Total RNA LT - (with Ribo-ZeroTM Human/Mouse/ Rat) - Set A |
Illumina |
RS-123–2201 |
TruSeq® Stranded Total RNA LT - (with Ribo-ZeroTM TM Human/ Mouse/Rat) - Set B |
Illumina |
RS-123–2202 |
High-Throughput Library Preparation Kit Standard PCR Amp Module - 96 rxn |
KAPA Biosystems |
KK8234 |
RNeasy Mini Kit |
Qiagen |
Cat# 74104 |
Vi-CELL Reagent Quad Pak |
Beckman Coulter |
Cat# 383198 |
AlphaScreen GST Detection Kit, 500 assay points |
Perkin Elmer |
Cat# 6760603C |
AlphaScreen Streptavidin Donor beads |
Perkin Elmer |
Cat# 6760002S |
RNeasy MinElute Cleanup Kit |
Qiagen |
Cat# 74204 |
Glutathione Superflow Agarose |
Thermo |
Cat# 25236 |
Dynabeads® MyOne™ Streptavidin C1 |
Thermo |
Cat# 65001 |
Streptavidin M280 beads |
Invitrogen |
Cat# 11206D |
Dynabeads Protein G |
Invitrogen |
Cat# 10003D |
Phase Lock Gel Heavy |
VWR |
Cat# 10847–802 |
Trizol Reagent |
Invitrogen |
Cat# 15596–018 |
Crystal violet solution |
Sigma-Aldrich |
Cat# HT90132–1L |
Dead Cell Apoptosis Kit with Annexin V Alexa Fluor® 488 & Propidium Iodide (PI) |
Thermo |
Cat# V13245 |
CellTiter-Glo® 2.0 Assay |
Promega |
Cat# G9242 |
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Deposited Data |
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Raw and analyzed data |
This paper |
GEO: GSE112608
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Human reference genome GRCh37/hg19 |
Genome Reference Consortium |
https://www.ncbi.nlm.nih.gov/assembly/GCF_000001405.13/ |
Drosophila reference genome BDGP Release 5/dm3 |
Genome Reference Consortium |
http://www.fruitfly.org/sequence/release3genomic.shtml |
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Experimental Models: Cell Lines |
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HCT-116, human, male |
ATCC |
ATCC Cat# CCL-247, RRID:CVCL_0291 |
J-Lat 6.3 clone (Jurkat cells), human, male |
NIH AIDS Program |
NIH-ARP Cat# 9846-446, RRID:CVCL_8280 |
HEK-293T, human, male |
ATCC |
ATCC Cat# CRL-3216, RRID:CVCL_0063 |
HEK293T Flag-AFF1 |
(Lin et al., 2010) |
N/A |
HEK293T Flag-AFF4 |
(Lin et al., 2010) |
N/A |
NCI-H2171 [H2171] (ATCC® CRL-5929™), human, male |
ATCC |
ATCC Cat# CRL-5929, RRID:CVCL_1536 |
SW 1271 [SW1271] (ATCC® CRL-2177™), human, male |
ATCC |
ATCC Cat# CRL-2177, RRID:CVCL_1716 |
FAST Pol II Mutant E1126G HEK293 cells, human, female |
(Fong et al., 2014) |
N/A |
WT Pol II Mutant HEK293 cells, human, female |
(Fong et al., 2014) |
N/A |
Slow Pol II Mutant R749H HEK293 cells, human, female |
(Fong et al., 2014) |
N/A |
MDA231-LM2, human, female |
(Wang et al., 2017) |
N/A |
S2 cells, Drosophila, male |
Invitrogen |
Cat# R690–07 |
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Experimental Models: Organisms/Strains |
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athymic nude mice, nu/nu |
Envigo |
Hsd:Athymic NudeFoxn1nu
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Oligonucleotides |
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Recombinant DNA |
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shRNA targeting sequence: AFF1 #1 GCCTCAAGTGAAGTTTGACAA |
Sigma-Aldrich |
Clone ID: TRCN0000021975 |
shRNA targeting sequence: AFF1 #2 TAGGTTGGGAAAGCCGAAATA |
Sigma-Aldrich |
Clone ID: TRCN0000330908 |
shRNA targeting sequence: AFF4 #1 GCACGACCGTGAGTCATATAA |
Sigma-Aldrich |
Clone ID: TRCN0000426769 |
shRNA targeting sequence: AFF4 #2 GCACCAGTCTAAATCTATGTT |
Sigma-Aldrich |
Clone ID: TRCN0000015825 |
shRNA targeting sequence: ELL2 #1 AACGCCAGAATTATAAGGATG |
This paper |
N/A |
shRNA targeting sequence: ELL2 #2 AAATGATCCCCTCAATGAAGT |
This paper |
N/A |
Lenti-sh1368 knockdown c-myc shRNA targeting sequence: GACGAGAACAGTTGAAACA |
Addgene |
Addgene plasmid # 29435 |
pGEX-2TK cyclin T1 (1–300) |
Addgene |
Addgene plasmid # P432 |
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Software and Algorithms |
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TopHat 2.1.0 |
(Kim et al., 2013) |
https://ccb.jhu.edu/software/tophat/index.shtml |
MACS 1.4.2 |
(Zhang et al., 2008) |
http://liulab.dfci.harvard.edu/MACS/ |
EdgeR 3.12.0 |
(Robinson et al., 2010) |
http://bioconductor.statistik.tu-dortmund.de/packages/2.11/bioc/html/edgeR.html |
Bowtie version 1.1.2 |
(Langmead et al., 2009) |
http://bowtiebio.sourceforge.net/index.shtml |
Trimmomatic 0.33 |
(Bolger et al., 2014) |
http://www.usadellab.org/cms/index.php?page=trimmomatic |
Cutadapt 1.14 |
(Martin, 2011) |
https://cutadapt.readthedocs.io/en/stable/guide.html |
Bedtools 2.17 |
(Quinlan and Hall, 2010) |
https://launchpad.net/ubuntu/+source/bedtools/2.17.0-1 |
Metascape |
(Tripathi et al., 2015) |
http://metascape.org/gp/index.html |
R 3.2.1 |
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https://www.rproject.org/ |
ZINC database |
(Irwin et al., 2012) |
http://zinc.docking.org |
Small-Molecule Drug Discovery Suite 2017–2 |
Schrödinger |
https://www.schrodinger.com/suites/small-molecule-drugdiscovery-suite |
Prism 7 |
GraphPad Software |
https://www.graphpad.com |
FlowJo |
FlowJo, LLC |
https://www.flowjo.com |
Dassault Systèmes BIOVIA |
Dassault Systèmes |
https://www.3ds.com/productsservices/biovia/ |
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Other |
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