Reagents and tools table
| Reagent/Resource | Reference or Source | Identifier or Catalog Number | |
|---|---|---|---|
| Experimental Models | |||
| Jag1Ndr (C3HeB/FeJ-Jag1Ndr/Ieg) | (Andersson et al, 2018) | EMMA ID - EM:13207 | |
| Rag1−/− | Jackson Laboratories | Jax: 002216 | |
| Recombinant DNA | |||
| n.a. | n.a. | n.a. | |
| Antibodies | |||
| Antibodies used in immunohistochemical or western blot analysis | |||
| Anti-mouse-HRP | Abcam | WB 1:20,000 | #ab205719 |
| Anti-rabbit-HRP | Abcam | WB 1:20,000 | #ab205718 |
| Donkey anti-Rat IgG (H + L) Highly Cross-Adsorbed (also Ms) Secondary Antibody, Alexa Fluor™ 647 | ThermoFisher | IF 1:250 (1:500 for the FOXP3, ECAD, LCK staining) | A78947 |
| Anti-rabbit-AlexaFluor-488 | Jackson ImmunoResearch | IF 1:250 | 711-545-152 |
| Anti-mouse-RhodamineRedX | Jackson ImmunoResearch | IF 1:250 | 715-295-150 |
| Anti-β-actin | Santa Cruz Biotechnology | WB 1:2000 | #sc-47778 |
| Lck | Kind gift from Dominik Filipp | WB/IF 1:1000 | (Veillette et al, 1988) |
| Tyr505Lck | Cell Signaling | WB 1:1000 | #2751 |
| aSMA - paraffine sections | Dako | IF 1:50 | M0851 |
| aSMA - cryosections | Sigma-Aldrich | IF 1:500 | A2547 |
| Collagen I - paraffine sections | Abcam | IF 1:100 | ab21286 |
| ECAD - cryosections | BD Biosciences | IF 1:100 | 610181 |
| FOXP3 - paraffine sections | Thermo Fisher | IF 1:500 | FJK-16s - # 14-5773-80 |
| FOXP3 - cryosections | BD | IF 1:500 | 562996 |
| CK19 - paraffine sections | DSHB | IF 1:250 | TROMA-III RRID: AB_2133570 |
| Antibodies used in E9.5 flow cytometry. | |||
| CD41 | Biolegend | 1:100 | #133903; MWReg30 |
| CD117 | Invitrogen | 1:50 | #17-1172-83; ACK2 |
| Viability dye | Thermo Fisher | 1:1000 | #L34957 |
| TER119 | Invitrogen | 1:100 | #35-5921-82; TER-119 |
| FcRγ | Biolegend | 1:50 | #101317; 93 |
| Antibodies used in 25-color flow cytometry. | |||
| CD45 | BD Biosciences | 1:100 | 30-F11 |
| CD31 | BD Biosciences | 1:100 | 390 |
| CD49a | BD Biosciences | 1:400 | Ha31/8 |
| TCR-beta | BD Biosciences | 1:100 | H57-597 |
| TER119 | BD Biosciences | 1:200 | TER-119 |
| CD11b | BD Biosciences | 1:200 | M1/70 |
| EpCAM | BD Biosciences | 1:100 | G8.8 |
| Ly6C | BD Biosciences | 1:200 | AL-21 |
| CD34 | Biolegend | 1:50 | MEC14.7 |
| Streptavidin | BD Biosciences | 1:400 | NA |
| Tim-4 | BD Biosciences | 1:100 | RMT4-54 |
| Jag1 | Biolegend | 1:100 | HMJ1-29 |
| CD3 | Biolegend | 1:200 | 17A2 |
| MHC-II | Biolegend | 1:400 | I-A/I-E |
| CD117 | Biolegend | 1:50 | 2B8 |
| Viability dye | Thermo Fisher | 1:100 | NA |
| CD4 | Biolegend | 1:400 | RM4-5 |
| Ly6G | BD Biosciences | 1:200 | 1A8 |
| CD19 | Biolegend | 1:200 | 6D5 |
| Siglec-F | BD Biosciences | 1:200 | E50-2440 |
| CD71 | BD Biosciences | 1:100 | C2 |
| CD64 | BD Biosciences | 1:100 | X54-5/71 |
| Nestin | Thermo Fisher | 1:400 | 307501 |
| NKp46 | Biolegend | 1:50 | 29A1.4 |
| F4/80 | Biolegend | 1:100 | BM8 |
| CD8 | Thermo Fisher | 1:400 | 53-6.7 |
| CD11c | Biolegend | 1:200 | N418 |
| TruStain FcX (CD16/32) | Biolegend | 1:100 | 93 |
| Brilliant Stain Buffer Plus | BD Biosciences | 1:5 | N/A |
| Antibodies used P30 stage and transfer experiments. | |||
| CD4 | Biolegend | 1:400 | BV785 |
| CD44 | Biolegend | 1:800 | BV711 |
| CD8 | Biolegend | 1:400 | Percp-Cy5.5 |
| TCR-B | Biolegend | 1:200 | APC-Cy7 |
| Viability dye 506 | Thermo Scientific | 1:1000 | # 65-0866-14 |
| Foxp3 | eBioscience | 1:400 | BV421 |
| Oligonucleotides and other sequence-based reagents | |||
| n.a. | n.a. | n.a. | |
| Chemicals, Enzymes and other reagents | |||
| RPMI | Sigma-Aldrich | #R8758 | |
| DSS | Sigma-Aldrich | #D8906 | |
| PFA | VWR | #20909.290 | |
| TrueView | Vector Laboratories | SP-8500-15 | |
| Fluorescent mounting medium | Dako | S3023 | |
| RIPA Lysis and Extraction Buffer | Thermo Scientific | #89900 | |
| Protease and phosphatase inhibitor | Thermo Scientific | #A32961 | |
| Pierce™ BCA Protein Assay Kit | Thermo Scientific | #23225 | |
| Amersham™ Hybond® PVDF membranes | Sigma-Aldrich | # GE10600021 | |
| Pierce™ ECL Western Blotting Substrate | Thermo Scientific | #32209 | |
| HBSS | Gibco | #14175095 | |
| Dispase | Gibco | #17105041 | |
| FCS | Gibco | #10270106 | |
| EDTA | Thermo Scientific | #AM9262 | |
| fixation buffer from eBioscience™ FOXP3/ Transcription Factor Staining Buffer Set | eBioscience | #00-5523-00 | |
| Dead Cell Removal Kit | Miltenyi | #130-090-101 | |
| DPBS -Mg2+,-Ca2+ | Gibco | #D8537 | |
| Liver digest medium | Gibco | #17703034 | |
| TrypLE Express | Gibco | #12604013 | |
| 0.5M EDTA pH 8.0 | Thermo Scientific | #R1021 | |
| BSA | Sigma-Aldrich | #A7906 | |
| collagenase D | Roche | #11088858001 | |
| dispase II | Gibco | #17105041 | |
| DNase I | Sigma-Aldrich | #4716728001 | |
| Percoll | Sigma-Aldrich | #GE17-0891-01 | |
| ChromiumTM Single Cell 3’ Library and Gel Bead Kit v2 | 10X Genomics | PN-120237 | |
| Bioanalyzer High Sensitivity DNA kit | Agilent | #5067-4626 | |
| Software | |||
| Zen | Carl Zeiss | V3.4 | |
| Fiji | https://github.com/fiji/fiji | v2.9.0 | |
| FlowJo software | BD | V10.7.1 | |
| Other | |||
| 40 μm cell strainer | BD | #352350 | |
| 96-well V-bottom plates | Sigma-Aldrich | #BR781601 | |
| 2 ml Eppendorf DNA LoBind® tubes | Merck | #EP0030108078 | |
| Millex-GV Filter 0.22 µm | Merck-Millipore | #SLGVR33RS | |
| EASYstrainer | Greiner | #542040 | |
| 70 μm filter cap | Falcon | #38030 | |