KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Anti-HaloTag® pAb | Promega | Cat#G9281 |
| Anti-RING1B mAb | MBL Life Science | Cat#D139–3 |
| Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 | Life Technologies | Cat#A-11029; RRID: AB_2534088 |
| Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 568 | Life Technologies | Cat#A11011; RRID: AB_143157 |
| Anti-PHC1 antibody (mAb) | Active Motif | Cat#39723; RRID: AB_2713961 |
| Bacterial and Virus Strains | ||
| DH5α | Life Technologies | Cat#18265–017 |
| Rosetta 2 (pLysS) host strains | Novagen | Cat#71403 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| ANTI-FLAG® M2 affinity gel | Millipore | Cat#A2220 |
| Doxycycline hyclate | Sigma Life Science | Cat#D9891 |
| Fetal Bovine Serum (FBS) | VWR | Cat#97068–085 |
| FLAG® Peptide | Sigma-Aldrich | Cat#F3290 |
| FluoroBrite DMEM Live-cell Imaging Medium | Life Technologies | Cat#A1896701 |
| GSH-Sepharose 4B beads | GE Healthcare | Cat#17–--01 |
| HaloTag® TMR Ligand | Promega | Cat#G8251 |
| IPTG | IBI Scientific | Cat#IB02105 |
| Janelia Fluor® 549 HaloTag® Ligand | Promega | Cat#GA1110 |
| Janelia Fluor® 646 HaloTag® Ligand | Promega | Cat#GA1120 |
| Leukemia Inhibitor Factor | Purified in lab | N/A |
| Polyethylene glycol (PEG) | Rigauku Reagents | Cat#1008062 |
| SYBR Gold | Invitrogen | Cat#S11494 |
| Critical Commercial Assays | ||
| Amicon Ultra Centrifugal Filter Units | Merck Millipore | Cat#UFC501024 |
| Bradford Assay | Thermo Scientific | Cat#1856209 |
| CoverWell perfusion chamber gasket | Thermo Scientific | Cat#C18139 |
| Novex 4–20% Tris-Glycine Protein gel | Life Technologies | Cat#EC6026BOX |
| NuPAGE 4–12% Bis-Tris Protein Gel | Invitrogen | Cat#NP0321BOX |
| Spectra/Por 1 Dialysis Membrane | Spectrum Labs | Cat#132645T |
| Experimental Models: Cell Lines | ||
| Human: HEK293T | Tatavosian et al., 2019 | N/A |
| Mouse: PGK12.1: Cbx2−/− | Katoh-Fukui et al., 1998 | N/A |
| Mouse: PGK12.1: Eed−/− | Endoh et al., 2008 | N/A |
| Mouse: PGK12.1: HT-CBX2/Eed−/− | Tatavosian et al., 2019 | N/A |
| Mouse: PGK12.1: HT-CBX2/Mel18−/−/Bm1i−/− | Tatavosian et al., 2019 | N/A |
| Mouse: PGK12.1: HT-CBX2 | Zhen et al., 2016 | N/A |
| Mouse: PGK12.1: HT-CBX2/Ring1a−/−/Ring1bfl/fl; Rosa26::CreERT2 | Tatavosian et al., 2019 | N/A |
| Mouse: PGK12.1: HT-CBX21−192 | This study | N/A |
| Mouse: PGK12.1: HT-CBX21−281 | This study | N/A |
| Mouse: PGK12.1: HT-CBX21−498 | This study | N/A |
| Mouse: PGK12.1: HT-CBX265−532 | This study | N/A |
| Mouse: PGK12.1: HT-CBX289−532 | This study | N/A |
| Mouse: PGK12.1: HT-CBX2ATLm | This study | N/A |
| Mouse: PGK12.1: HT-CBX2ATL-P2A | This study | N/A |
| Mouse: PGK12.1: HT-CBX2ATm | This study | N/A |
| Mouse: PGK12.1: HT-CBX2AT-P2A | This study | N/A |
| Mouse: PGK12.1: HT-CBX2CA-P2A | This study | N/A |
| Mouse: PGK12.1: HT-CBX2cd | This study | N/A |
| Mouse: PGK12.1: HT-CBX2hncr-P2A | This study | N/A |
| Mouse: PGK12.1: HT-CBX2HPCR-P2A | This study | N/A |
| Mouse: PGK12.1: HT-CBX2SRR-S2A | This study | N/A |
| Mouse: PGK12.1: HT-CBX2SRR-S2E | This study | N/A |
| Mouse: PGK12.1: HT-CBX4 | Zhen et al., 2016 | N/A |
| Mouse: PGK12.1: HT-CBX6 | Zhen et al., 2016 | N/A |
| Mouse: PGK12.1: HT-CBX7 | Zhen et al., 2016 | N/A |
| Mouse: PGK12.1: HT-CBX8 | Zhen et al., 2016 | N/A |
| Mouse: PGK12.1: HT-MEL18 | This study | N/A |
| Mouse: PGK12.1: HT-NLS | Zhen et al., 2016 | N/A |
| Mouse: PGK12.1: HT-PHC1 | This study | N/A |
| Mouse: PGK12.1: HT-RING1B | This study | N/A |
| Mouse: Mel18−/−/Bmi1−/− | Elderkin et al., 2007 | N/A |
| Mouse: PGK12.1 | Penny et al., 1996 | N/A |
| Mouse: Ring1a−/−/Ring1bfl/fl; Rosa26::CreERT2 | Endoh et al., 2008 | N/A |
| Oligonucleotides | ||
| Major Satellite DNA: | IDT | N/A |
| 5′-GGACCTGGAATATGGCG | ||
| AGAAAACTGAAAATCACGG | ||
| AAAATGAGAAATACACACTTTAG-3′ | ||
| AT-mutated satellite DNA: | IDT | N/A |
| 5′-GGACCTGGAGTCTGGCG | ||
| AGAGAGCTGAGACTCACGG | ||
| ACACTGAGAGACACACACCTGAG-3′ | ||
| Recombinant DNA | ||
| Plasmid: pGEX-6P-1-GST-CBX2AT-P2A-FLAG | This study | N/A |
| Plasmid: pGEX-6P-1-GST-CBX2-FLAG | Tatavosian et al., 2019 | N/A |
| Plasmid: pMD2.G | Zhen et al., 2016 | N/A |
| Plasmid: pSPAX2 | Zhen et al., 2016 | N/A |
| Plasmid: pTRIPZ (M1)-CERULEAN-MEL18 | Zhen et al., 2016 | N/A |
| Plasmid: pTRIPZ (M1)-CERULEAN-PHC1 | Zhen et al., 2016 | N/A |
| Plasmid: pTRIPZ (M1)-HT-CBX2 | Zhen et al., 2016 | N/A |
| Plasmid: pTRIPZ (M1)-HT-CBX21−192 | This study | N/A |
| Plasmid: pTRIPZ (M1)-HT-CBX21−281 | This study | N/A |
| Plasmid: pTRIPZ (M1)-HT-CBX21−498 | This study | N/A |
| Plasmid: pTRIPZ (M1)-HT-CBX265−532 | This study | N/A |
| Plasmid: pTRIPZ (M1)-HT-CBX289−532 | This study | N/A |
| Plasmid: pTRIPZ (M1)-HT-CBX2ATLm | This study | N/A |
| Plasmid: pTRIPZ (M1)-HT-CBX2ATL-P2A | This study | N/A |
| Plasmid: pTRIPZ (M1)-HT-CBX2ATm | This study | N/A |
| Plasmid: pTRIPZ (M1)-HT-CBX2AT-P2A | This study | N/A |
| Plasmid: pTRIPZ (M1)-HT-CBX2CA-P2A | This study | N/A |
| Plasmid: pTRIPZ (M1)-HT-CBX2cd | This study | N/A |
| Plasmid: pTRIPZ (M1)-HT-CBX2hncr-P2A | This study | N/A |
| Plasmid: pTRIPZ (M1)-HT-CBX2HPCR-P2A | This study | N/A |
| Plasmid: pTRIPZ (M1)-HT-CBX2SRR-S2A | This study | N/A |
| Plasmid: pTRIPZ (M1)-HT-CBX2SRR-S2E | This study | N/A |
| Plasmid: pTRIPZ (M1)-HT-MEL18 | This study | N/A |
| Plasmid: pTRIPZ (M1)-HT-PHC1 | This study | N/A |
| Plasmid: pTRIPZ (M1)-HT-RING1B | This study | N/A |
| Plasmid: pTRIPZ (M1)-YFP-RING1B | Zhen et al., 2014 | N/A |
| Protein: pGEX-6P-1-GST-CBX2-FLAG | This study | N/A |
| Protein: pGEX-6P-1-GST-CBX2AT-P2A-FLAG | This study | N/A |
| Software and Algorithms | ||
| Adobe Illustrator | Adobe Inc | N/A |
| Adobe Photoshop | Adobe Inc | N/A |
| ImageJ | NIH | https://imagej.nih.gov/ |
| MATLAB | MathWorks | N/A |
| OriginLab | OriginLab Corporation | N/A |
| SlideBook 6.0 Software | 3i | N/A |
| Spot-On | Hansen et al., 2018 | N/A |
| U-track | Jaqaman et al., 2008 | https://github.com/DanuserLab/u-track |
| Other | ||
| Alpha Pan-Apochromat 100 × /1.40 Oil immersion Objective | Zeiss, Germany | N/A |
| Alpha Pan-Apochromat 100 × /1.46 Oil immersion Objective | Zeiss, Germany | N/A |
| Axio Observer D1 Microscope | Zeiss, Germany | N/A |
| Brightline® single-band laser filter set (Semrock; excitation filter: FF02–482/18–25, emission filter: FF01–525/45–25, and dichroic mirror: Di02-R488–25 × 36) | Semrock | N/A |
| Brightline® single-band laser filter set (Semrock; excitation filter: FF01–561/14, emission filter: FF01–609/54, and dichroic mirror: Di02-R561–25 × 36) | Semrock | N/A |
| Brightline® single-band laser filter set (Semrock; excitation filter: BLP01–635R-25, emission filter: FF01–640/14–25, and dichroic mirror: Di02-R635–25 × 36) | Semrock | N/A |
| ChemiDoc XRS+ System | Bio-Rad | N/A |
| Evolve 512 × 512 EMCCD camera | Photometrics; Tuscon, AZ | Cat#1708265 |
| Single Channel Temperature Controller | Warner Instruments | Cat#TC-324 |
| Solid-State LaserStack | 3i | N/A |
| Sonicator | Vibra-cell | VCX130 |