KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| 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 |
| Bacterial and Yeast Strains | ||
| Rosetta 2(DE3) competent bacterial cells | Novagen | Cat# 71402 |
| S. cerevisiae BY4741 strain | Euroscarf | Cat# Y00000 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |