KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER | |
|---|---|---|---|
| Antibodies | |||
| CD31 | BD Pharmingen | 557355; RRID: AB_396660 | |
| pSMAD1/5/9 | Cell Signaling | 41D10; RRID:AB_491015 | |
| Laminin | Sigma | L9393; RRID: AB_477163 | |
| PDGFRA | R&D Systems | AF1062; RRID: AB_2236897 | |
| Alpha-smooth muscle actin (ACTA2) | Abcam | 5694; RRID: AB_2223021 | |
| CD81 | Invitrogen | 13–0811-81; RRID: AB 466514 | |
| GFP | Abcam | 6556/6662; RRIDs: AB 305564 and AB 305635 | |
| Lysozyme | Dako | A0099; RRID: AB_2341230 | |
| CD45 | eBiosciences | 17–0451-82; RRID:AB 469392 | |
| EPCAM | BioLegend | 118214; RRID:AB 1134102 | |
| Chemicals, Peptides, and Recombinant Proteins | |||
| 0.25%Trypsin-EDTA | Gibco | 25200–056 | |
| Collagenase II | Worthington | LS004176 | |
| RSPONDIN3 (HUMAN) | R&D Systems | 3500-RS | |
| RSPONDIN2 (HUMAN) | R&D Systems | 3266-RS | |
| RSPONDIN3 | R&D Systems | 4120-RS | |
| RSPONDIN2 | R&D Systems | 6946-RS | |
| GREM2 | R&D Systems | 2069-PR | |
| BMP2 | Peprotech | 120–02 | |
| BMP7 | R&D Systems | 5666-BP | |
| NOGGIN | Peprotech | 120–10C | |
| RSPONDIN1 (HUMAN) | R&D Systems | 4645-RS | |
| EGF | ThermoFisher | PHG0311 | |
| GREMLIN1 | ThermoFisher | 956GR050 | |
| Matrigel | Corning | 356231 | |
| Diphtheria toxin | Enzo Life Sciences | BML-G135–0001 | |
| 4-OH Tamoxifen | Sigma | T5648 | |
| DMEM/F12 tissue culture medium | Gibco | 12634–010 | |
| Fetal bovine serum | Corning | 35–010-CV | |
| Penicillin/Streptomycin | Life Technologies | 15140163 | |
| Glutamax | Gibco | 35050–661 | |
| N-2 Supplement (100X) | Life Technologies | 17502001 | |
| B-27 Supplement (50X) | Life Technologies | 17504001 | |
| NICOTINAMIDE | Sigma | 72340 | |
| N-Acetylcysteine | Sigma | A9165 | |
| Normal goat serum | Cell Signaling | 5425S | |
| DAPI | BD Pharmingen | 5649097 | |
| Dulbecco-s modified Eagle medium (DMEM) | Corning | 17–205-CV | |
| Hanks’ balanced salt solution (HBSS, 10X) | Gibco | 14065–056 | |
| Phosphate buffered saline (PBS, 10X) | Corning | 46–013-CM | |
| AlexaFluor goat 546 anti-rat IgG | Invitrogen | A11081 | |
| AlexaFluor goat 633 anti-rabbit IgG | Invitrogen | A21071 | |
| AlexaFluor 594 donkey anti-goat IgG | Invitrogen | A11058 | |
| OCT compound | Tissue-Tek | 4583 | |
| Biotin | Invitrogen | 17–4317-82 | |
| Bovine serum albumin | Sigma | A9647 | |
| Paraformaldehyde | EMS | 15714-S | |
| Picric acid | Fluka | 80456 | |
| FocusClear | CElExplorer | FC-101 | |
| Triton-X 100 | Sigma | T8787 | |
| Sodium azide | Sigma | S2002 | |
| HEPES | Sigma | H3375 | |
| VectaShield mounting medium | Vector Laboratories | H-1000 | |
| 5-bromo-2-deoxyuridine | Life Technologies | B23151 | |
| Power SYBR | Thermofisher | 4367659 | |
| Phalloidin | Invitrogen | A12381 | |
| Critical Commercial Assays | |||
| RNAscope Multiplex Fluorescent Reagent Kit v2 | Advanced Cell Diagnostics (ACD) | 323100 | |
| Single Cell 3’ V3.1 assay | 10X Genomics | PN-1000147 | |
| Deposited Data | |||
| Raw and analyzed data | This paper; Table S2 | GEO: GSE184158 | |
| Mouse reference genome Mm10, GRCm38 | Genome Reference Consortium | https://www.ncbi.nlm.nih.gov/grc/mouse | |
| Adult mesenchymal RNA datasets | McCarthy, et al.30 | GEO: GSE130681 | |
| Experimental Models: Organisms/Strains | |||
| Mouse: PdgfraH2BeGFP | Jackson Laboratory | 007669 | |
| Mouse: ROSA26LSL-TdTomato | Jackson Laboratory | 007909 | |
| Mouse: Myh11Cre(ER-T2) | Jackson Laboratory | 019079 | |
| Mouse: Etv1Cre(ER-T2) | Jackson Laboratory | 013048 | |
| Mouse: PdgfraCre(ER-T2> | Jackson Laboratory | 032770 | |
| Mouse: ROSA26L-S-L-DTA | Jackson Laboratory | 009669 | |
| Oligonucleotides | |||
| Grem1 probes | Advanced Cell Diagnostics (ACD) | 314741 | |
| Grem2 probes | ACD | 473981 | |
| Hhip probes | ACD | 448441 | |
| Noggin probes | ACD | 467391 | |
| Olfm4 probes | ACD | 311831 | |
| Osr1 probes | ACD | 496281 | |
| Axin2 probes | ACD | 400331 | |
| Rspo3 probes | ACD | 483781 | |
| Rspo2 probes | ACD | 402008 | |
| Rspo1 probes | ACD | 479591 | |
| Id1 probes | ACD | 312221 | |
| Bmp5 probes | ACD | 401241 | |
| Pcp4 probes | ACD | 402311 | |
| Id1 | GAGGCGGCATGTGTTCCA | CTGGAGGCTGAAAGGTGGAG | |
| Id2 | AAAAACAGCCTGTCGGACCA | GGTGATGCAGGCTGACGATA | |
| Vil1 | GGGATCCCTTCAAGTGGAGTA | TGGTATTGGCAGT GAAAA CG | |
| Alpi | CAGCCCTTTCCAATTTCACTG | AGGACATCGCCACTCAACTC | |
| Lgr5 | CCTACTCGAAGACTTACCCAGT | GCATTGGGGTGAATGATAGCA | |
| Cd81 | ACACCTTCTACGTGGGCATC | TGCTTCACATCCTTGGCGAT | |
| Bmp5 | TAGATGTGGGCTGGCTTGTC | ACCTCGCTTGCCTTGAAGAA | |
| Grem1 | CTGGAGACCCAGAGTACCGT | CGGTGCGATTCATTCTGTCAC | |
| Rspo3 | ATGTGGCTTCAAAAGGGGGA | TGGGGTCTCAATGCTGGACT | |
| Olfm4 | CAGCCACTTTCCAATTTCACTG | GCTGGACATACTCCTTCACCTTA | |
| Software and Algorithms | |||
| GraphPad Prism v9.0.2 | GraphPad | ||
| R | R | https://cran.r-proiect.org/bin/windows/ | |
| Seurat package v4.1 | Butler et al.73 | https://github.com/satiialab/seurat | |
| Cell Ranger v3.1.1 | 10X Genomics | https://support.10xgenomics.com/single-cell-gene-expression/software/pipelines/latest/installation | |
| BioMart v2.38.0 | Durinck et al.74 | https://github.com/grimbough/biomaRt | |
| MetaNeighbor | Crow et al.58 | https://github.com/maggiecrow/MetaNeighbor | |
| ImageJ Fiji | Schindelin et al.75 | https://imagei.net/Fiii/ | |
| Biorender | BioRender.com | ||
| FlowJo Software v10 | FlowJo | ||
| Other | |||
| ACD HybEZ II Hybridization System | ACD | 321710 | |
| 10X Genomics Controller | 10X Genomics | ||
| EdU labeling kit | Invitrogen | C10640 | |
| Slide spacers | Grace Bio-Labs | 654002 | |