KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Histone H3 antibody | Abcam | ab1791 |
| HA antibody | Abcam | ab9110 |
| Flag | Sigma | F3165 |
| Flag-M2 beads | Sigma | A2220 |
| Sox2 | R&D Systems | MAB2018 |
| Klf4 | R&D Systems | AF3158 |
| Actin | Sigma | A5441 |
| Bacterial and Virus Strains | ||
| Biological Samples | ||
| Chemicals, Peptides, and Recombinant Proteins | ||
| Doxycycline | Sigma | D9891–5G |
| Poly-L-ornithine | Sigma | P4957–50ML |
| U2AF2 | Guo et al., 2019 | N/A |
| HNRNPA1 | Guo et al., 2019 | N/A |
| SRSF2 | Guo et al., 2019 | N/A |
| NANOG | This study | N/A |
| RARA | This study | N/A |
| CTCF | This study | N/A |
| OCT4 | This study | N/A |
| MYC | This study | N/A |
| P53 | This study | N/A |
| KLF4 | This study | N/A |
| ESR1 | This study | N/A |
| YY1 | This study | N/A |
| SOX2 | This study | N/A |
| STAT3 | This study | N/A |
| GATA2 | This study | N/A |
| SMAD3 | This study | N/A |
| KLF4-ΔARM (aa 355–386) | This study | N/A |
| SOX2-ΔARM (aa 118–178) | This study | N/A |
| GATA2-ΔARM (aa 360–395) | This study | N/A |
| GFP | This study | N/A |
| BamHI | NEB | R0136 |
| Critical Commercial Assays | ||
| Dual Luciferase Assay Kit | Promega | E1960 |
| NEBuilder HiFi DNA Assembly Master Mix | NEB | E2621S |
| Monarch Gel Extraction Kit | NEB | T1020S |
| Phusion polymerase | NEB | M0531S |
| MEGAscript T7 Transcription Kit | Invitrogen | AM1334 |
| MEGAclear Transcription Clean-Up Kit | Invitrogen | AM1908 |
| Cy5-labeled UTP | Enzo LifeSciences | ENZ-42506 |
| Lipofectamine 3000 Transfection Agent | ThermoFisher | L3000001 |
| CUT&Tag-IT Assay Kit | Active Motif | 53160 |
| Deposited Data | ||
| RBR-ID mass spectrometry proteomics data | This study | ProteomeXchange: PXD035484 |
| Original imaging data and single molecule traces | This study | Mendeley Data DOI: 10.17632/dkx9gsh42 h.1 |
| GATA2 K562 CLIP-seq | This study | GEO GSE232181 |
| SOX2 and KLF4 CUT&Tag sequencing | This study | GEO GSE232181 |
| Experimental Models: Cell Lines | ||
| V6.5 murine embryonic stem cells | Jaenisch laboratory | N/A |
| pJH308_PBFH_NtermHalo_hsKLF4_WT mESCs | This study | N/A |
| pJH309_PBFH_NtermHalo_hsKLF4_ΔARM (aa 355–386) mESCs | This study | N/A |
| pJH294_PBFH_NtermHalo_hsSOX2_WT mESCs | This study | N/A |
| pJH295_PBFH_NtermHalo_hsSOX2_ΔARM (aa 118–178) mESCs | This study | N/A |
| pJH290_PBFH_NtermHalo_hsCTCF_WT mESCs | This study | N/A |
| pJH291_PBFH_NtermHalo_hsCTCF_ΔARM (aa 576–611) mESCs | This study | N/A |
| pJH357_PBFH-NtermHalo-hsGATA2-WT K562 cells | This study | N/A |
| pJH358_PBFH-NtermHalo-hsGATA2_ΔARM (aa 360–395) K562 cells | This study | N/A |
| pJH337_PBFH-NtermHalo-RUNX1-WT K562 cells | This study | N/A |
| pJH342_PBFH-NtermHalo-RUNX1-_ΔARM (aa 174–197) K562 cells | This study | N/A |
| DNA, RNA and peptides sequences | ||
| See Table S4 for all peptide, RNA, gblock and oligonucleotide sequences | This study | N/A |
| Recombinant DNA | ||
| pJH201_CBFH_NtermGFP_hsNANOG_WT | This study | N/A |
| pJH203_CBFH_NtermGFP_hsRARA_WT | This study | N/A |
| pJH205_CBFH_NtermGFP_hsCTCF_WT | This study | N/A |
| pJH199_CBFH_NtermGFP_hsOCT4_WT | This study | N/A |
| pJH200_CBFH_NtermGFP_hsMYC_WT | This study | N/A |
| pJH204_CBFH_NtermGFP_hsP53_WT | This study | N/A |
| pJH278_CBFH_CtermGFP_hsKLF4_WT | This study | N/A |
| pJH202_CBFH_NtermGFP_hsESR1_WT | This study | N/A |
| pJH087_CBFH_NtermGFP_hsYY1_WT | This study | N/A |
| pJH198_CBFH_NtermGFP_hsSOX2_WT | This study | N/A |
| pJH227_CBFH_NtermGFP_hsSTAT3_WT | This study | N/A |
| pJH247_CBFH_NtermGFP_hsGATA2_WT | This study | N/A |
| pJH226_CBFH_NtermGFP_hsSMAD3_WT | This study | N/A |
| pJH279_CBFH_CtermGFP_hsKLF4_ΔARM (aa 355–386) | This study | N/A |
| pJH245_CBFH_NtermGFP_hsSOX2_ΔARM (aa 118–178) | This study | N/A |
| pJH272_CBFH_NtermGFP_hsGATA2_ΔARM (aa 360–395) | This study | N/A |
| pJH308_PBFH_NtermHalo_hsKLF4_WT | This study | N/A |
| pJH309_PBFH_NtermHalo_hsKLF4_ΔARM (aa 355–386) | This study | N/A |
| pJH294_PBFH_NtermHalo_hsSOX2_WT | This study | N/A |
| pJH295_PBFH_NtermHalo_hsSOX2_ΔARM (aa 118–178) | This study | N/A |
| pJH290_PBFH_NtermHalo_hsCTCF_WT | This study | N/A |
| pJH291_PBFH_NtermHalo_hsCTCF_ΔARM (aa 576–611) | This study | N/A |
| pJH357_PBFH-NtermHalo-hsGATA2-WT | This study | N/A |
| pJH358_PBFH-NtermHalo-hsGATA2_ΔARM (aa 360–395) | This study | N/A |
| pJH337_PBFH-NtermHalo-RUNX1-WT | This study | N/A |
| pJH342_PBFH-NtermHalo-RUNX1-_ΔARM (aa 174–197) | This study | N/A |
| pJH325_HIV-LTR-LUC | This study | N/A |
| pJH326_HIV-LTR-ΔTAR-LUC | This study | N/A |
| pJH327_pcDNA3-HIV-tat-WT | This study | N/A |
| pJH329_pcDNA3-HIV-tat-ARM>KLF4-BP | This study | N/A |
| pJH330_pcDNA3-HIV-tat-ARM>SOX2-BP | This study | N/A |
| pJH361_pcDNA3-HIV-tat-ARM-R/KtoA | This study | N/A |
| pJH371_pcDNA3-HIV-tat-ARM-to-GATA2-peptide | This study | N/A |
| pJH365_pcDNA3-HIV-tat-ARM-to-ESR1 | This study | N/A |
| pJH366_pcDNA3-HIV-tat-ARM-to-ESR1-R269C | This study | N/A |
| pJH438_pb-hUbiC-Gal4-KLF4-WT | This study | N/A |
| pJH439_pb-hUbiC-Gal4-KLF4-ARM-KRtoA | This study | N/A |
| pJH441_pb-hUbiC-Gal4-KLF4-SWAP-Tat-ARM | This study | N/A |
| pJH375_pb-hUbiC-tetR-KLF4-WT | This study | N/A |
| pJH376_pb-hUbiC-tetR-KLF4-ARM-KRtoA | This study | N/A |
| pJH377_pb-hUbiC-tetR-KLF4-SWAP-Tat-ARM | This study | N/A |
| pJH437_pJP080_UAS_luciferase | Li et al., 2020 | N/A |
| pJH175_4xTet_Luc_reporter | This study | N/A |
| Software and Algorithms | ||
| Fiji image processing package | Schindelin et al., 2012 | https://fiji.sc/ |
| Prism | GraphPad | https://www.graphpad.com/scientific-software/prism/ |
| Code generated by the study | This study | Zenodo DOI: 10.5281/zenodo.797 4933 https://zenodo.org/record/7974933 |
| Other | ||
| 35 mm glass-bottom imaging dishes | Mattek Corporation | P35G-1.5–20-C |