Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference |
Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (Mus musculus) | Ebf1+/- | Pongubala et al., 2008 | Provided by Dr. Grosschedl, Max Planck Institute of Immunobiology and Epigenetics |
|
| Genetic reagent (Mus musculus) | B6.Cg-Tg(BCL2)25Wehi/J | Jackson Laboratory | Stock# 002320 | |
| Genetic reagent (Mus musculus) | B6.Gt(ROSA)26Sortm1.1(CAG-cas9*,-EGFP)Fezh/J | Jackson Laboratory | Stock# 024858 | |
| Cell line (Mus musculus) | OP9 | Yokoyama et al., 2013 | Stromal cell line derived from fetal murine calvaria (B6 x C3H, op/op) | |
| Cell line (Mus musculus) | TD7 | This paper | Mouse fetal thymus (B6, E15.5)-derived mesenchymal cell line | |
| Cell line (Mus musculus) | Pro-B(+) | This paper | Ebf1-deficient fetal liver-derived hematopoietic progenitor cell line | |
| Cell line (Mus musculus) | Pro-B(−) | This paper | Ebf1-deficient fetal liver-derived hematopoietic progenitor cell line | |
| Cell line (Mus musculus) | OP9-Dll4 | Hirano et al., 2020 | ||
| Cell line (Homo sapiens) | HEK293T | Hirano et al., 2020 | RRID:CVCL_0063 | |
| Cell line (Homo sapiens) | PLAT-E | Hirano et al., 2020 | RRID:CVCL_B488 | |
| Antibody | FITC anti-mouse CD4 (Rat monoclonal) | BD Biosciences | Cat# 561835 RRID:AB_10894386 | FC (1:500) |
| Antibody | PE anti-mouse CD4 (Rat monoclonal) | BD Biosciences | Cat# 561829 RRID:AB_10926205 | FC (1:500) |
| Antibody | APC anti-mouse CD8a (Rat monoclonal) | BioLegend | Cat# 100711 RRID:AB_312750 | FC (1:500) |
| Antibody | APCCy7 anti-mouse CD8a (Rat monoclonal) | BioLegend | Cat# 100713 RRID:AB_312752 | FC (1:500) |
| Antibody | PerCpCy5.5 anti-mouse CD11b (Rat monoclonal) | BioLegend | Cat# 101227 RRID:AB_893233 | FC (1:500) |
| Antibody | PECy7 anti-mouse CD19 (Rat monoclonal) | Tonbo Biosciences | Cat# 60-0193 RRID:AB_2621840 | FC (1:250) |
| Antibody | PerCpCy5.5 anti-mouse CD25 (Rat monoclonal) | eBioscience | Cat# 45-0251-82 RRID:AB_914324 | FC (1:1000) |
| Antibody | APC-e780 anti-mouse CD25 (Rat monoclonal) | eBioscience | Cat# 47-0251-82 RRID:AB_1272179 | FC (1:200) |
| Antibody | FITC anti-mouse CD44 (Rat monoclonal) | BioLegend | Cat# 103005 RRID:AB_312956 | FC (1:500) |
| Antibody | APCCy7 anti-mouse CD44 (Rat monoclonal) | BioLegend | Cat# 103027 RRID:AB_830784 | FC (1:500) |
| Antibody | PECy7 anti-mouse CD45 (Rat monoclonal) | eBioscience | Cat# 25-0451-82 RRID:AB_2734986 | FC (1:400) |
| Antibody | APC anti-mouse CD45 (Rat monoclonal) | BioLegend | Cat# 103111 RRID:AB_312976 | FC (1:1000) |
| Antibody | PE anti-mouse CD45.1 (Mouse monoclonal) | BioLegend | Cat# 110727 RRID:AB_893348 | FC (1:250) |
| Antibody | PerCpCy5.5 anti-mouse CD45.2 (Mouse monoclonal) | BioLegend | Cat# 109807 RRID:AB_313444 | FC (1:250) |
| Antibody | PerCpCy5.5 anti-mouse B220 (Rat monoclonal) | BioLegend | Cat# 103235 RRID:AB_893356 | FC (1:200) |
| Antibody | APCCy7 anti-mouse Thy1.2 (Rat monoclonal) | BioLegend | Cat# 105327 RRID:AB_10613280 | FC (1:1000) |
| Antibody | FITC anti-mouse Gr-1 (Rat monoclonal) | BioLegend | Cat# 108405 RRID:AB_313370 |
FC (1:500) |
| Antibody | Biotin anti-mouse Gr-1 (Rat monoclonal) | eBioscience | Cat# 13-5931-86 RRID:AB_466802 | FC (1:300) |
| Antibody | Biotin anti-mouse TER119 (Rat monoclonal) | eBioscience | Cat# 13-5921-85 RRID:AB_466798 | FC (1:300) |
| Antibody | Biotin anti-mouse CD11b (Rat monoclonal) | eBioscience | Cat# 13-0112-86 RRID:AB_466361 | FC (1:300) |
| Antibody | Biotin anti-mouse CD11c (Armenian Hamster monoclonal) | eBioscience | Cat# 13-0114-85 RRID:AB_466364 | FC (1:300) |
| Antibody | Biotin anti-mouse CD19 (Rat monoclonal) | eBioscience | Cat# 13-0193-85 RRID:AB_657658 | FC (1:300) |
| Antibody | Biotin anti-mouse NK1.1 (Rat monoclonal) | eBioscience | Cat# 13-5941-85 RRID:AB_466805 | FC (1:300) |
| Antibody | Biotin anti-mouse CD3ε (Armenian Hamster monoclonal) | eBioscience | Cat# 13-0031-82 RRID:AB_466319 | FC (1:300) |
| Antibody | PE anti-rat CD2 (Mouse monoclonal) | BioLegend | Cat# 201305 RRID:AB_2073811 | FC (1:500) |
| Antibody | PE anti-human NGFR (Mouse monoclonal) | eBioscience | Cat# 12-9400-42 RRID:AB_2572710 | FC (1:500) |
| Antibody | PE Hamster IgG (Armenian Hamster monoclonal) |
BioLegend | Cat# 400907 RRID:AB_326593 | FC (1:200) |
| Antibody | PE anti-mouse Notch1 (Armenian Hamster monoclonal) |
BioLegend | Cat# 130607 RRID:AB_1227719 | FC (1:200) |
| Antibody | PE anti-mouse Notch2 (Armenian Hamster monoclonal) |
BioLegend | Cat# 130707 RRID:AB_1227725 | FC (1:200) |
| Antibody | PE anti-mouse Notch3 (Armenian Hamster monoclonal) |
BioLegend | Cat# 130507 RRID:AB_1227733 | FC (1:200) |
| Antibody | PE anti-mouse Notch4 (Armenian Hamster monoclonal) |
BioLegend | Cat# 128407 RRID:AB_1133997 | FC (1:200) |
| Antibody | Alexa Fluor 647 Mouse IgG1 (Mouse monoclonal) | BioLegend | Cat# 400130 RRID:AB_2800436 | FC (2 µl per test) |
| Antibody | Alexa Fluor 647 anti-GATA3 (Mouse monoclonal) | BD Biosciences | Cat# 560068 RRID:AB_1645316 | FC (8 µl per test) |
| Antibody | Rabbit-IgG (Rabbit monoclonal) | Cell Signaling Technology | Cat# 3900 RRID:AB_1550038 | FC (1:500) |
| Antibody | Anti-TCF1 (Rabbit monoclonal) | Cell Signaling Technology | Cat# 2203 RRID:AB_2199302 | FC (1:100) |
| Antibody | Anti-LMO2 (Rabbit monoclonal) | Abcam | Cat# ab91652 RRID:AB_2049879 | FC (1:200) ChIP (2ug/sample) |
| Antibody | Anti-c-Myc (Rabbit monoclonal) | Cell Signaling Technology | Cat# 5605 RRID:AB_1903938 | FC (1:100) |
| Antibody | DyLight 488 anti-rabbit IgG (Donkey polyclonal) |
BioLegend | Cat# 406404 RRID:AB_1575130 | FC (1:250) |
| Antibody | DyLight 649 anti-rabbit IgG (Donkey polyclonal) |
BioLegend | Cat# 406406 RRID:AB_1575135 | FC (1:500) |
| Antibody | Anti-Tubulinα (Mouse monoclonal) | Sigma | Cat# T6199 RRID:AB_477583 | WB (1:1000) |
| Antibody | Anti-LMO2 (Mouse monoclonal) | Novus | Cat# NB110-78626 RRID:AB_1084895 | WB (1:1000) ChIP (2 µg/sample) |
| Antibody | Anti-human LMO2 (Goat polyclonal) | R&D Systems | Cat# AF2726 RRID:AB_2249968 | ChIP (2 µg/sample) |
| Antibody | Anti-H3K4me3 (Rabbit polyclonal) | Millipore | Cat# 07-473 RRID:AB_1977252 | ChIP (5 µl/sample) |
| Recombinant DNA reagent (plasmid) | pCMV-VSV-G-RSV-Rev | RIKEN BRC | Cat# RDB04393 | Lentiviral packaging plasmid |
| Recombinant DNA reagent (plasmid) | pCAG-HIVgp | RIKEN BRC | Cat# RDB04394 | Lentiviral packaging plasmid |
| Recombinant DNA reagent (plasmid) | pLVS-EF-IR2 | This paper | Lentiviral vector with IRES-rat CD2 | |
| Recombinant DNA reagent (plasmid) | mLmo2/pLVS-EF-IR2 | This paper | pLVS-EF-IR2 Lentiviral vector encoding mLmo2 |
|
| Recombinant DNA reagent (plasmid) | mTcf7/pLVS-EF-IR2 | This paper | pLVS-EF-IR2 Lentiviral vector encoding mTcf7 |
|
| Recombinant DNA reagent (plasmid) | GCDN | Hirano et al., 2015 | Retroviral vector with IRES-human NGFR | |
| Recombinant DNA reagent (plasmid) | hBCL2/GCDN | This paper | GCDN Retroviral vector encoding hBCL2 |
|
| Recombinant DNA reagent (plasmid) | mMeis1/GCDN | This paper | GCDN Retroviral vector encoding mMeis1 |
|
| Recombinant DNA reagent (plasmid) | mHmga2/GCDN | This paper | GCDN Retroviral vector encoding mHmga2 |
|
| Recombinant DNA reagent (plasmid) | Cas9-GFP | Hosokawa et al., 2018 | Retroviral vector to express Cas9 and GFP |
|
| Recombinant DNA reagent (plasmid) | E42-dTet | Hosokawa et al., 2018 | Retroviral vector to express sgRNA and human NGFR |
|
| Recombinant DNA reagent (plasmid) | sgRNA against Luciferase (control) | Hosokawa et al., 2018 | 5’-ggcatttcgcag cctaccg-3’ |
|
| Recombinant DNA reagent (plasmid) | sgRNA against LMO2 #1 | This paper | 5’-tcgatggccgag gacattg-3’ |
|
| Recombinant DNA reagent (plasmid) | sgRNA against LMO2 #2 | This paper | 5’-aatgtcctcggc catcgaa-3’ |
|
| Recombinant DNA reagent (plasmid) | sgRNA against LMO2 #3 | This paper | 5’-gaaagccatcga ccagtac-3’ |
|
| Sequence-based reagent | ActB (Forward) | This paper | PCR primers | 5’-tacagcccgggg agcat-3’ |
| Sequence-based reagent | ActB (Reverse) | This paper | PCR primers | 5’-acacccgccac cagttc-3’ |
| Sequence-based reagent | Meis1 (Forward) | This paper | PCR primers | 5’-gacgctttaaag agagataaagatgc-3’ |
| Sequence-based reagent | Meis1 (Reverse) | This paper | PCR primers | 5’- catttctcaaa aatcagtgctaaga -3’ |
| Sequence-based reagent | Hmga2 (Forward) | This paper | PCR primers | 5’-aaggcagcaaaa acaagagc-3’ |
| Sequence-based reagent | Hmga2 (Reverse) | This paper | PCR primers | 5’-gccgtttttctc caatggt-3’ |
| Sequence-based reagent | Bcl11a (Forward) | This paper | PCR primers | 5’-ccaaacaggaac acacatagcaga-3’ |
| Sequence-based reagent | Bcl11a (Reverse) | This paper | PCR primers | 5’-ggggattagagc tccgtgt-3’ |
| Sequence-based reagent | Gata3 (Forward) | This paper | PCR primers | 5’-ttatcaagccca agcgaag-3’ |
| Sequence-based reagent | Gata3 (Reverse) | This paper | PCR primers | 5’-tggtggtggtct gacagttc-3’ |
| Sequence-based reagent | Lmo2 (Forward) | This paper | PCR primers | 5’-gaggcgcctcta ctacaa-3’ |
| Sequence-based reagent | Lmo2 (Reverse) | This paper | PCR primers | 5’-gatccgcttgtcacaggatg-3’ |
| Sequence-based reagent | Tcf7 (Forward) | This paper | PCR primers | 5’-cagctcccccatactgtgag-3’ |
| Sequence-based reagent | Tcf7 (Reverse) | This paper | PCR primers | 5’-tgctgtctatatccgcaggaa-3’ |
| Sequence-based reagent | Tcf7 promoter region (Forward) | This paper | PCR primers | 5’-ttaagtttttattggtgaatgagtt-3’ |
| Sequence-based reagent | Tcf7 promoter region (Reverse) | This paper | PCR primers | 5’-aaaaaactccaaaaataaaacccac-3’ |
| Sequence-based reagent | Tcf7 TSS (Forward) | This paper | PCR primers | 5’-gcagcaagggttgcattt-3’ |
| Sequence-based reagent | Tcf7 TSS (Reverse) | This paper | PCR primers | 5’-ttgtctgtactgggctgtttacat-3’ |
| Sequence-based reagent | Tcf7 -31kb (Forward) | This paper | PCR primers | 5’-ttccatccaccgttttgttt-3’ |
| Sequence-based reagent | Tcf7 -31kb (Reverse) | This paper | PCR primers | 5’-ggcgtgtggtgggaatacta-3’ |
| Sequence-based reagent | Tcf7 -35kb (Forward) | This paper | PCR primers | 5’-ctgcaagcagctggaagtc-3’ |
| Sequence-based reagent | Tcf7 -35kb (Reverse) | This paper | PCR primers | 5’-cactggaagctgtgagtgatg-3’ |
| Sequence-based reagent | Igk 3’UTR (Forward) | This paper | PCR primers | 5’-ggcacatctgttgctttcgc -3’ |
| Sequence-based reagent | Igk 3’UTR (Reverse) | This paper | PCR primers | 5’-ggggtaggga gcaggtgtat-3’ |
| Peptide, recombinant protein | PerCpCy5.5 streptavidin | BioLegend | Cat# 405214 | FC (1:200) |
| Peptide, recombinant protein | Recombinant Mouse SCF | PeproTech | Cat# 250-03 | |
| Peptide, recombinant protein | Recombinant Human FLT3L | PeproTech | Cat# 300-19 | |
| Peptide, recombinant protein | Recombinant Mouse IL-7 | PeproTech | Cat# 217-17 | |
| Commercial assay or kit | Foxp3 / Transcription Factor Staining Set | eBioscience | Cat# 00-5523-00 | Used to detect TCF1 and GATA3 |
| Commercial assay or kit | Fixation/Permeabilization Solution Kit with BD GolgiStop | BD Biosciences | Cat# 554715 | Used to detect Lmo2 and c-Myc |
| Commercial assay or kit | Permeabilization Buffer Plus | BD Biosciences | Cat# 561651 | Used to detect Lmo2 and c-Myc |
| Commercial assay or kit | High Capacity cDNA Reverse Transcription Kit | Thermo Fisher Scientific | Cat# 4368814 | |
| Commercial assay or kit | NucleoSpin Tissue | TaKaRa Bio | Cat# 740952.50 | |
| Commercial assay or kit | MethylEasy Xceed | TaKaRa Bio | Cat# ME002 | |
| Commercial assay or kit | TaKaRa EpiTaq HS (for bisulfite-treated DNA) | TaKaRa Bio | Cat# R110A | |
| Commercial assay or kit | Mighty TA-cloning kit | TaKaRa Bio | Cat# 6028 | |
| Commercial assay or kit | Anti-Biotin MicroBeads | Miltenyi Biotec | Cat# 130-090-485 | |
| Commercial assay or kit | Dynabeads Protein A | Thermo Fisher Scientific | Cat# 10001D | |
| Commercial assay or kit | Dynabeads Protein G | Thermo Fisher Scientific | Cat# 10003D | |
| Commercial assay or kit | Dynabeads M-280 Sheep Anti-Rabbit IgG | Thermo Fisher Scientific | Cat# 11203D | |
| Commercial assay or kit | NE-PER Nuclear and Cytoplasmic Extraction Reagents | Pierce | Cat# 78833 | |
| Commercial assay or kit | PCR purification Kit | Qiagen | Cat# 28004 | |
| Commercial assay or kit | Fast SYBR Green Master Mix | Thermo Fisher Scientific | Cat# 4385614 | |
| Commercial assay or kit | NEBNext Ultra II DNA Library Prep with Sample Purification Beads | NEB | Cat# E7103S | |
| Commercial assay or kit | NEBNext Multiplex Oligos for Illumina | NEB | Cat# E7500S | |
| Chemical compound, drug | Trizol reagent | Thermo Fisher Scientific | Cat# 15596026 | |
| Chemical compound, drug | 7-AAD | BioLegend | Cat# 420403 | FC (1:50) |
| Chemical compound, drug | Alexa Fluor 647 Annexin V | BioLegend | Cat# 640911 | FC (1:40) |
| Chemical compound, drug | DSG (disuccinimidyl glutarate) | Thermo Fisher Scientific | Cat# 20593 | 1 mg/ml |
| Software, algorithm | FlowJo | BD Biosciences | RRID:SCR_008520 |