KEY RESOURCE TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Cd34 | Thermo Fisher Scientific | 14-0341-82 |
| Dpp4 | R&D | AF954-SP |
| EBF-2 | R&D | AF7006 |
| Gata6 | RnD | AF1700-SP |
| Gata6 | Cell Signaling Technology | 5851T |
| GFP | SantaCruz | SC-101536 |
| GFP | Abcam | AB13970 |
| Keratin 14 | Biolegend | 906001 |
| mCherry | OriGene | AB0040-200 |
| mCherry | Neuromics | CH22115 |
| MTC02 | Abcam | AB3298 |
| Myh11 | Alfa Aesar | J64817 |
| MyHCfast | Sigma-Aldrich | M4276 |
| Perilipin 1 | Progen | GP29 |
| Perilipin-1 | Abcam | AB61682 |
| PPARG | Thermo Fisher Scientific | MA5-14889 |
| RFP | Rockland | 600-901-379 |
| RFP | Rockland | 600-401-379 |
| Sca-1/Ly6 | R&D | AF1226-SP |
| TBX6 | Abcam | AB38883 |
| UCP1 | R&D | MAB6158 |
| Donkey Anti-Rat IgG H&L (Alexa Fluor® 647) preadsorbed | Abcam | A-11015 |
| Donkey anti-Goat IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate | Thermo Fisher Scientific | A21206 |
| Donkey anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 647 | Thermo Fisher Scientific | A-21207 |
| Donkey anti-Sheep IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 594 | Thermo Fisher Scientific | A-31573 |
| Donkey anti-Sheep IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate | Life Technologies | A-31572 |
| Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 | Thermo Fisher Scientific | 703-545-155 |
| Donkey anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor® 594 | Thermo Fisher Scientific | 703-165-155 |
| Donkey anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor® 647 | Thermo Fisher Scientific | A-31571 |
| Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 555 | Thermo Fisher Scientific | 706-545-148 |
| Alexa Fluor® 488 AffiniPure Donkey Anti-Chicken IgY (IgG) (H+L) | Jackson ImmunoResearch Laboratories | 703-545-155 |
| Cy™3 AffiniPure Donkey Anti-Chicken IgY (IgG) (H+L) | Jackson ImmunoResearch Laboratories | 703-165-155 |
| Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 647 | Thermo Fisher Scientific | A-31571 |
| Alexa Fluor® 488 AffiniPure Donkey Anti-Guinea Pig IgG (H+L) | Jackson ImmunoResearch Laboratories | 706-545-148 |
| Bacterial and virus strains | ||
| Promega JM109 Competent E.Coli Cells | Thermo Fisher Scientific | PR-L1001 |
| Biological samples | ||
| Human fetal tissues | University of Washington Birth Defects Research Laboratory (BDRL) under a protocol approved by the University of Washington Institutional Review Board | 98 days (H28540), 115 days (H28572), 122 days (H28560), 125 days (H28626) |
| Chemicals, peptides, and recombinant proteins | ||
| Y-27632 dihydrochloride | R&D Systems | 1254/10 |
| CHIR 99021 | R&D Systems | 4423 |
| LDN-193189 | Stemgent | 04-0074 |
| FGF-2 | PeproTech | 450-33 |
| HGF | PeproTech | 315-23 |
| IGF-1 | PeproTech | 250-19 |
| TGFb1 | PeproTech | 100-21-10 |
| BMP7 | Thermo Fisher Scientific | PHC9544 |
| 3,3′,5-Triiodo-L-thyronine sodium salt | Sigma-Aldrich | T6397 |
| Porcn Inhibitor II C59 | Millipore Sigma | 500496 |
| SB431542 | Selleck Chemicals | S1067 |
| Dexamethasone | Sigma-Aldrich | D4902 |
| Hydrocortisone | Sigma-Aldrich | H0888 |
| Rosiglitazone | Sigma-Aldrich | R2408 |
| EGF | PeproTech | AF-100-15 |
| 3-Isobutyl-1-methylxanthine | Sigma-Aldrich | I7018 |
| ERthermAC | Sigma | SCT057 |
| Oligomycin | Tocris | 4110 |
| Forskolin | Sigma-Aldrich | F6886 |
| Antimycin A | Sigma-Aldrich | A8674 |
| Rotenone | Sigma-Aldrich | R8875 |
| 4% Paraformaldehyde | Electron Microscopy Sciences | 15710 |
| Critical commercial assays | ||
| NucleoSpin® RNA kit | Macherey and Nagel | 740955 |
| iScript™ cDNA Synthesis Kit | Bio-Rad | 1708891 |
| iTaq™ Universal SYBR® Green Supermix | Bio-Rad | 172-5124 |
| Deposited data | ||
| Single cell RNA seq data | This paper | GEO (GSE185623) |
| Bulk RNA seq data | This paper | GEO (GSE185518) |
| Experimental models: Cell lines | ||
| NCRM-1 (NIH CRM control iPSC line (male)) human iPSC | RUCDR Infinite Biologics | ND50028 |
| NCRM-1-UCP1-H2B-mCherry - 2 clones | This paper | N.A. |
| NCRM-1-PAX3-NLS-Venus - 2 clones | This paper | N.A. |
| Experimental models: Organisms/strains | ||
| Mouse : Pax7-iCre/+;Rosa26-mTmG/+ | This study; Gabrielle Kardon Lab | N.A. |
| Mouse : Gata6-EGFPCreERT2;Rosa26-fl/STOP/fl-tdTomato | This study; Fiona Watt Lab | N.A. |
| Mouse : wildtype CD1 IGS mice | Charles River | 022 |
| Oligonucleotides | ||
| RT-qPCR primers | This study | Table S2 |
| Recombinant DNA | ||
| PSpCas9 (BB)-2A-GFP(PX458) | Addgene | 48138 |
| pUC19 vector backbone | New England Biolabs | E5510S |
| pUC19-5’HA-H2B-mCherry-P2A-3’HA | This study | N.A. |
| Software and algorithms | ||
| ImageJ | Image J | https://imagej.nih.gov/ij/ |
| Prism 9 | GraphPad | https://www.graphpad.com |
| Single cell analysis code | This study | https://doi.org/10.5281/zenodo.8071624 |
| kNN-classifier | Diaz-Cuadros et al., 23 | N.A. |
| Stream | Chen et al., 25 | N.A. |
| Waddington-OT kernel | Schiebinger et al., 26 | N.A. |
| ProFAT webtool | Cheng et al., 37 | N.A. |
| FastQC (v0.11.9) and used STAR RNA-seq aligner (v2.7.3a) | Dobin et al., 61 | N.A. |
| featureCounts (v2.0.1) | Liao et al., 62 | N.A. |
| DESeq2 package (v 1.22.2) | Love et al., 63 | N.A. |
| EnrichR | Kuleshov et al., 64 | N.A. |
| Scrublet | Wolock et al., 65 | N.A. |
| Scanpy (v.1.6.0) | Wolf et al., 70 | N.A. |
| Cellrank | Lange et al., 71 | N.A. |