Antibodies |
GAPDH Loading Control Monoclonal Antibody (GA1R) |
Invitrogen™ |
Cat# MA5–15738 |
Monoclonal ANTI-FLAG® M2 antibody |
Sigma-Aldrich |
Cat# F1804 |
AEBP2 (D7C6X) Rabbit mAb |
Cell Signaling |
Cat# 14129S |
Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb |
Cell Signaling |
Cat# 9733 |
EZH2 (D2C9) XP® Rabbit mAb |
Cell Signaling |
Cat# 5246S |
SUZ12 (D39F6) XP® Rabbit mAb |
Cell Signaling |
Cat# 3737S |
Monoclonal Anti-HA antibody produced in mouse |
Sigma-Aldrich |
Cat# H9658 |
Myc-Tag antibody (mAb) |
Active motif |
Cat# 39279 |
MTF2 Antibody |
Protein tech |
Cat# 16208–1-AP |
PHF19 Antibody |
Cell Signaling |
Cat# 77271S |
HA-Tag (C29F4) Rabbit mAb |
Cell Signaling |
Cat# 3724S |
RBBP4 Antibody |
Bethyl Laboratories |
Cat# A301–206A |
Anti-EED Antibody, clone AA19 |
EMD Millipore |
Cat# 05–1320 |
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Bacterial and Yeast Strains |
Rosetta 2(DE3) competent bacterial cells |
Novagen |
Cat# 71402 |
S. cerevisiae BY4741 strain |
Euroscarf |
Cat# Y00000 |
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Chemicals, Peptides, and Recombinant Proteins |
Dynabeads™ Protein A for Immunoprecipitation |
Invitrogen™ |
Cat# 10002D |
Dynabeads™ Protein G for Immunoprecipitation |
Invitrogen™ |
Cat# 10004D |
Pierce™ 16% Formaldehyde (w/v), Methanol-free |
Thermo Scientific™ |
Cat# 28908 |
cOmplete, EDTA-free proteinase inhibitor cocktail |
Sigma-Aldrich |
Cat# 11873580001 |
RNase A |
Thermo Scientific™ |
Cat# EN0531 |
Proteinase K, recombinant, PCR Grade |
Sigma-Aldrich |
Cat# 3115836001 |
Glycogen |
Invitrogen™ |
Cat# 10814010 |
SYBR™ Green PCR Master Mix |
Applied Biosystems™ |
Cat# 4309155 |
Gelatin solution-100 mL Type B, 2% in H2O |
Sigma-Aldrich |
Cat# G-1393 |
DMEM/F12 (1:1) |
Gibco™ |
Cat# 11320033 |
Neurobasal Medium |
Gibco™ |
Cat# 21103049 |
Gibco™ KnockOut™ DMEM |
Gibco™ |
Cat# 10829018 |
GlutaMAX™ Supplement |
Gibco™ |
Cat# 35050061 |
BSA Fraction V 7.5% |
Gibco™ |
Cat# 15260037 |
MEM Non-essential Amino Acid Solution (100×) |
Sigma-Aldrich |
Cat# M7145–100ML |
2-Mercaptoethanol |
Sigma-Aldrich |
Cat# M3148–25ML |
N-2 supplement |
Gibco™ |
Cat# 17502048 |
B27 supplement |
Gibco™ |
Cat# 17504044 |
PD0325901-MEK inhibitor |
Cayman |
Cat# 391210109 |
CHIR99021 (GSK3 inhibitor) |
Cayman |
Cat# NC0226336 |
TrypLE™ Express Enzyme (1X), phenol red |
Gibco™ |
Cat# 12605028 |
Penicillin-Streptomycin |
Sigma-Aldrich |
Cat# P0781–100ML |
HyClone™ Fetal Bovine Serum, USDA Tested |
GE Healthcare |
Cat# SH3091003 |
Dulbecco’s Modified Eagle’s Medium - high glucose |
Sigma-Aldrich |
Cat# D6429–500ML |
Opti-MEM-Reduced Serum Medium |
Gibco™ |
Cat# 31985062 |
Xtreme Gene 9 DNA transfection |
Roche |
Cat# 6365787001 |
Lenti-X™ Concentrator |
Clontech |
Cat# 631232 |
Gibco™ Sterile Puromycin Dihydrochloride |
Gibco™ |
Cat# A1113803 |
IgG Sepharose™ 6 Fast Flow Agarose |
GE Healthcare |
Cat# 17096901 |
Strep-Tactin® Superflow® high capacity 50% suspension |
IBA-Lifesciences |
Cat# 21208010 |
Pierce™ Anti-DYKDDDDK Magnetic Agarose |
Thermo Scientific™ |
Cat# A36797 |
Pierce™ Avidin Agarose |
Thermo Scientific™ |
Cat# 20219 |
Pierce™ Glutathione Agarose |
Thermo Scientific™ |
Cat# 16101 |
HIS-Select® Nickel Affinity Gel |
Sigma-Aldrich |
Cat# P6611–25ML |
d-Desthiobiotin |
Sigma-Aldrich |
Cat# D1411–1G |
SuperSignal™ West Pico PLUS Chemiluminescent Substrate |
Thermo Scientific™ |
Cat# 34580 |
SYBR Gold |
ThermoFisher |
Cat# S11494 |
IPTG |
Gold Biotechnology |
Cat# I2481C |
PfuUltra High-Fidelity DNA Polymerase |
Agilent Technologies |
Cat# 600385 |
ProtoScript® II First Strand cDNA Synthesis Kit |
NEB |
Cat# E6560S |
Bio-Rad gel filtration standard |
BioRad |
cat # 1511901 |
Yeast tRNA (10 mg/mL) |
Invitrogen |
cat # AM7119 |
Amersham Hybond-N+ |
GE Healthcare |
cat # RPN203B |
EMD Millipore™ Novagen™ Mouse Genomic DNA |
EMD Millipore |
Cat# 69239–3 |
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Deposited Data |
Crystal structure of SUZ12(N)-RBBP4-PHF19(RC)-JARID2(TR) |
This study |
PDB: 6NQ3 |
Uncropped gel images |
This study |
http://dx.doi.org/10.17632/j8mddywf7j.1 |
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Experimental Models: Cell Lines |
HEK293T |
ATCC |
Cat# CRL-3216 |
SUZ12-KO HEK293T |
This study |
N/A |
SUZ12-KO + SUZ12WT HEK293T |
This study |
N/A |
SUZ12-KO + SUZ123D HEK293T |
This study |
N/A |
SUZ12-KO mESCs |
Kristian Helin Laboratory |
N/A |
SUZ12-KO + SUZ12WT mESCs |
This study |
N/A |
SUZ12-KO + SUZ123D mESCs |
This study |
N/A |
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Recombinant DNA |
pCS2-Myc-EZH2 |
This paper |
N/A |
pCS2-FLAG-EZH2 |
This paper |
N/A |
pCS2-FLAG-EZH2(H689A) |
This paper |
N/A |
pCS2-His6-EED |
This paper |
N/A |
pCS2-HA-RBBP4 |
This paper |
N/A |
pCS2-ProteinA-TEV-HA-SUZ12WT
|
This paper |
N/A |
pCS2-ProteinA-TEV-HA-SUZ123D
|
This paper |
N/A |
pCS2-ProteinA-TEV-HA-Suz12R196A
|
This paper |
N/A |
pCS2-ProteinA-TEV-HA-Suz12W334A
|
This paper |
N/A |
pCS2-ProteinA-TEV-HA-Suz12G518W
|
This paper |
N/A |
pCS2-Flag-PHF19WT
|
This paper |
N/A |
pCS2-Flag-PHF19ΔDS(531−544)
|
This paper |
N/A |
pCS2-Flag-PHF19K331A/K332A
|
This paper |
N/A |
pCS2-Flag-MTF2WT
|
This paper |
N/A |
pCS2-Flag-MTF2ΔDS(544−557)
|
This paper |
N/A |
pCS2-Flag-MTF2K338A/K339A
|
This paper |
N/A |
pET-28a-His6-SUMO-StrepII-Flag-MTF2(1–593) |
This paper |
N/A |
pGEX-4T1-TEV-PHF19(500–580) |
This paper |
N/A |
pGEX-4T1-TEV-MTF2(507–593) |
This paper |
N/A |
pET-28a-His6-SUMO-PHF1(493–567) |
This paper |
N/A |
pGEX-4T1-TEV-AEBP2(407–503) |
This paper |
N/A |
pFastBac-Flag-Ezh2(1–746) |
This paper |
N/A |
pFastBac-His6-Thrombin-RBBP4(1–425) |
This paper |
N/A |
pACEBac1-His6-Flag-EED(1–441)-3C-StrepII |
This paper |
N/A |
pACEBac1-His6-Flag-SUZ12(1–739)-TEV-StrepII |
This paper |
N/A |
pACEBac1-StrepII-3C-AEBP2(1–295)-His6
|
This paper |
N/A |
pACEBac1-StrepII-AEBP2(1–301) |
This paper |
N/A |
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Software and Algorithms |
Pymol |
The PyMOL Molecular Graphics System, Schrö dinger, LLC. |
N/A |
PHENIX |
https://www.phenix-online.org |
N/A |
Coot |
https://www2.mrc-lmb.cam.ac.uk/personal/pemsley/coot/ |
N/A |