Skip to main content
. 2017 Jul 20;67(2):181–193.e5. doi: 10.1016/j.molcel.2017.06.009
REAGENT or RESOURCE SOURCE IDENTIFIER
Antibodies

α-UBR5 Abcam Cat#ab70311; RRID: AB_2210186
α-GFP (rabbit) Sigma Cat#G1544; RRID: AB_439690
α-GFP (mouse) Sigma Cat#G6539; RRID: AB_259941
α-Flag (mouse) Sigma Cat#F1804; RRID: AB_262044
α-Flag (rabbit) Sigma Cat#F7425; RRID: AB_439687
α-HA (rat) Sigma Cat#3F10
α-HA (rabbit) Abcam Cat#ab9110; RRID: AB_307019
α-Myc Santa Cruz Biotechnology Cat#sc-789; RRID: AB_631274
α-active β-catenin (ABC) Cell Signaling Technologies Cat#8814S; RRID: AB_11127203
α-β-catenin BD Transduction Laboratories Cat#610153
α-XIAP BD Transduction Laboratories Cat#610763; RRID: AB_398086
α-HUWE1 Abcam Cat#ab70161; RRID: AB_1209511
α-TRIP12 Abcam Cat#ab86220; RRID: AB_1925533
α-HECTD1 Abcam Cat#ab101992; RRID: AB_10711075
α-UBE3C Abcam Cat#ab101512; RRID: AB_10711205
α-β-tubulin Sigma Cat#T4026; RRID: AB_477577
α-TLE1-4 Santa Cruz Biotechnology Cat#sc-13373; RRID: AB_2203721
α-GST Abcam Cat#ab19256; RRID: AB_444809
HRP conjugated Goat α-Mouse Santa Cruz Biotechnology Cat#sc-2005; RRID: AB_631736
HRP conjugated Goat α-Rabbit Santa Cruz Biotechnology Cat#sc-2301; RRID: AB_650500
HRP conjugated Goat α-Rat Santa Cruz Biotechnology Cat#sc-2032; RRID: AB_631755
HRP conjugated Donkey?α-Goat R&D Systems Cat#HAF109; RRID: AB_357236
Alexa Fluor 488 conjugated Goat α-Rabbit Life Technologies Cat#A11008
Alexa Fluor 488 conjugated Goat α-Mouse Life Technologies Cat#A11029
Alexa Fluor 546 conjugated Goat α-Mouse Life Technologies Cat#A11003
Alexa Fluor 647 conjugated Goat α-Guinea pig Invitrogen Cat#A21450
α-Senseless Prof. Hugo J. Bellen N/A
α-Vestigial Prof. Sean B. Carroll N/A
α-Wingless DSHB Cat#4D4; RRID: AB_528512
α-Armadillo DSHB Cat#N27A1; RRID: AB_528089

Chemicals, Peptides, and Recombinant Proteins

Ni-NTA Agarose QIAGEN Cat#30210
α-FLAG M2 Affinity Gel Sigma Cat#A2220
EZview Red α-HA Affinity Gel Sigma Cat#E6779
GFP-trap_A Chromotek Cat#gta-20
Glutathione Sepharose 4b GE Healthcare Cat#17075601
Dynabeads MyOne Streptavidin C1 Invitrogen Cat#65001
Lipofectamine2000 Invitrogen Cat#11668019
Polyethylenimine, linear, MW25000 Polysciences Cat#23966
EDTA-free Protease Inhibitor Cocktail Roche Cat#04693159001
MG132 Sigma Cat#C2211
NMS-873 Cayman Chemical co. Cat#1418013-75-8
CB-5083 Cayman Chemical co. Cat#1542705-92-9
Cycloheximide Sigma Cat#C104450
Puromycin dihydrochloride Sigma Cat#P8833
3xFLAG-Peptide Sigma Cat#F4799
L-Glutathione reduced Sigma Cat#G4251
VectaShield with DAPI Vector Laboratories Cat#H-1200
Ubiquitin-activating enzyme E1 (UBE1A) Boston Biochem Cat#E-305
Ubiquitin-conjugating enzyme E2 (UBE2L3) Boston Biochem Cat#E2-640
Ubiquitin Boston Biochem Cat#U-100
Methyl-ubiquitin Boston Biochem Cat#U-501
K6-only ubiquitin Boston Biochem Cat#UM-K60
K11-only ubiquitin Boston Biochem Cat#UM-K110
K27-only ubiquitin Boston Biochem Cat#UM-K270
K29-only ubiquitin Boston Biochem Cat#UM-K290
K33-only ubiquitin Boston Biochem Cat#UM-K330
K48-only ubiquitin Boston Biochem Cat#UM-K480
K63-only ubiquitin Boston Biochem Cat#UM-K630
Histone H2A (1-22) - GK(Biotin) AnaSpec Cat#64639-1
Histone H3 (1-21) Biotinylated AnaSpec Cat#AS-61702
Histone H4 (1-23) - GGK(Biotin) AnaSpec Cat#AS-65097
KOD DNA polymerase Merck Millipore Cat#71086-4
Phusion DNA polymerase NEB Cat#M0530L

Critical Commercial Assays

Dual-Luciferase Reporter Assay System Promega Cat#E1910
UbiCREST Deubiquitinase Enzyme Kit Boston Biochem Cat#K-400
RNeasy Mini Kit (RNA Purification) QIAGEN Cat#74104
iScript cDNA synthesis kit Biorad Cat#170-8890
SYBR Select Master Mix Applied Biosystems Cat#4472908

Deposited Data

Raw Imaging Data This paper http://dx.doi.org/10.17632/j2wjdkj5cn.1

Experimental Models: Cell Lines

HEK293T ATCC Cat#CRL-3216
HeLa ATCC Cat#CCL-2
HCT116 ATCC Cat#CCL-247

Experimental Models: Organisms/Strains

D. melanogaster: hydK7-19 Prof. Jessica E. Treisman FlyBase: FBal0144234
D. melanogaster: groMB36 Prof. David Ish-Horowicz FlyBase: FBal0230454
D. melanogaster: axinP Prof. Tetsu Akiyama FlyBase: FBal0097414
D. melanogaster: UAS.SoxF Dr. Fernando Casares FlyBase: FBtp0051564
D. melanogaster: pygoS123 Bloomington Drosophila Stock Center FlyBase: FBal0146872
Bloomington: 7209
D. melanogaster: UAS.ArmS10 Bloomington Drosophila Stock Center FlyBase: FBtp0001723
Bloomington: 4782
D. melanogaster: UAS.MamDN Prof. Sarah Bray FlyBase: FBtp0014588
D. melanogaster: Vg-Gal4, UAS-flp; FRT82b GFP Bienz Laboratory Miller et al., 2013

Oligonucleotides

Primer sequences for RT-qPCR, see Table S3 This paper N/A

Recombinant DNA

Plasmid: pCMV-tag2b-Flag-UBR5 Prof. Rina Rosin-Arbesfeld Hay-Koren et al., 2011
Plasmid: pCS2 Myc-TLE3 Prof. Ethan Lee Hanson et al., 2012
Plasmid: pcDNA3.1 VCP-GFP Prof. Nico Dantuma Tresse et al., 2010
Plasmid: pcDNA3.1 HA-Δ45-β-catenin Bienz Laboratory Morin et al., 1997
Plasmid: pET GST-HES1 Prof. Stefano Stifani Grbavec and Stifani, 1996
Plasmid: pCMV His-ubiquitin Dr. Thomas Mund Mund et al., 2015
Plasmid: pcDNA3.1 HA-PAIP2 Bienz Laboratory N/A
Plasmid: pcDNA3.1 SSDP-GFP Bienz Laboratory Fiedler et al., 2015
Plasmid: pcDNA3.1 LDB1-GFP Bienz Labaratory Fiedler et al., 2015
Plasmid: pcDNA3.1 GFP-TCF4 Bienz Labaratory N/A
Plasmid: pcDNA3.1 GFP-Pygo1 Bienz Labaratory N/A
Plasmid: pSpCas9(BB)-2A-GFP (PX458) Addgene Ran et al., 2013; Addgene#48138
Plasmid: pSpCas9(BB)-2A-Puro (PX459) Addgene Ran et al., 2013; Addgene#62988
Plasmid: pTA-Luc m50 Super 8x TopFLASH Addgene Veeman et al., 2003; Addgene#12456
Plasmid: pRL-CMV Renilla luciferase Promega Cat#E2261