KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Chicken anti-GFP | Abcam | Cat# ab13970; RRID: AB_300798 |
| Donkey anti-chicken IRDye 680RD | LI-COR Biosciences | Cat#: 926-68075; RRID:AB_10974977 |
| Donkey anti-mouse IRDye 680RD | LI-COR Biosciences | Cat#: 926-68072; RRID: AB_10953628 |
| Goat anti-guinea pig Alexa 594 | Thermo Fisher | Cat# A-11076; RRID: AB_141930 |
| Goat anti-mouse Alexa 488 | Thermo Fisher | Cat# A-11001; RRID: AB_2534069 |
| Goat anti-rabbit IRDye 800CW | LI-COR Biosciences | Cat#: 925-32211; RRID: AB_2651127 |
| Guinea pig anti-PLIN1 | Fitzgerald | Cat# 20R-PP004; RRID: AB_1288416 |
| Mouse anti-TFRC (H68.4) | Invitrogen | Cat# 13-6890; RRID: AB_86624 |
| Mouse anti-TOM20 (F10) | Santa Cruz Biotechnology | Cat# sc-17764; RRID: AB_628381 |
| Rabbit Anti-b-actin (13E5) | Cell Signaling Technology | Cat# 4970; RRID: AB_2223172 |
| Rabbit anti-Ferritin-H | Cell Signaling Technology | Cat# 4393; RRID: AB_11217441 |
| Rabbit anti-Fpn1 | Novus Biologicals | Cat# NBP1-21502; RRID: AB_2302075 |
| Rabbit anti-HIF-2a | Novus Biologicals | Cat# NB100-122SS; RRID: AB_10000872 |
| Rabbit anti-MGAT2 | Proteintech | Cat# 19514-1-AP; RRID: AB_10638917 |
| Chemicals, peptides, and recombinant proteins | ||
| 3a70B complete counting cocktail | RPI | SKU: 111154 |
| 3H-Triolein | PerkinElmer | Cat#: NEC075H050UC |
| 14C-Palmiate | PerkinElmer | Cat#: NET431001MC |
| Acetaminophen | Sigma-Aldrich | Cat#: A7085 |
| ADMEM/F12 | Gibco | Cat#: 12634010 |
| Antifade mountant with DAPI | Invitrogen | Cat#: P36941 |
| AP20187 (Dimerizer) | Ariad Pharmaceuticals | N/A |
| Bathophenanthrolinedisulfonic acid disodium salt hydrate | Sigma-Aldrich | Cat#: B1375 |
| Benzonase | Sigma-Aldrich | Cat#: E8263 |
| BSA | Sigma-Aldrich | Cat#: A8806 |
| Collagenase IV | Roche | Cat#: 11088882001 |
| Dexamethasone | Sigma-Aldrich | Cat#: D4902 |
| Dispase II | Roche | Cat#: 04942078001 |
| DMEM high glucose | Gibco | Cat#: 11965118 |
| DMEM/F12 | Gibco | Cat#: 11320082 |
| Fetal Bovine Serum | Sigma-Aldrich | Cat#: 12303C |
| Glucose | Gibco | Cat#: 15023021 |
| GlutaMax | Gibco | Cat#: 35050079 |
| Glutaraldehyde solution | Sigma-Aldrich | Cat#: G5882 |
| Growth Factor-Reduced Matrigel | Corning | Cat#: 356230 |
| HEPES | Gibco | Cat#: 15630080 |
| Insulin | Eli Lilly | Product ID: A10008415 |
| Intralipid | Sigma-Aldrich | Cat#: 6I141 |
| Iron Standard for AAS | Sigma-Aldrich | Cat#: 16596 |
| Isobutylmethylxanthine | Sigma-Aldrich | Cat#: I7018 |
| Oil Red O | Sigma-Aldrich | Cat#: O0625 |
| Palmitate | Sigma-Aldrich | Cat#: P5585 |
| Paraformaldehyde | Sigma-Aldrich | Cat#: P6148 |
| PEI linear MW25000 | Polysciences | Cat#: 23966-100 |
| Penicillin and streptomycin | Gibco | Cat#: 15140122 |
| Phenol red free DMEM/F12 | Gibco | Cat#: 21041025 |
| Pluronic F-68 | ThermoFisher | Cat#: 24040032 |
| Potassium ferrocyanide | Sigma-Aldrich | Cat#: 60279 |
| Prolong Anti-Fade mounting medium | Invitrogen | Cat#: P36941 |
| RNase I | ThermoFisher | Cat#: EN0601 |
| Rosiglitazone | Sigma-Aldrich | Cat#: R2408 |
| Sarkosyl | Sigma-Aldrich | Cat#: L9150 |
| Sodium acetate | Sigma-Aldrich | Cat#: S2889 |
| Sodium cacodylate | Sigma-Aldrich | Cat#: C0250 |
| Thioglycolic acid solution | Sigma-Aldrich | Cat#: T6750 |
| Trizol | Invitrogen | Cat#: 12034977 |
| Tyloxapol | Sigma-Aldrich | Cat#: T0307 |
| Critical commercial assays | ||
| DAB Quanto chromogen and substrate kit | ThermoFisher | Cat#: TA-125-QHDX |
| HR Series NEFA-HR(2) Color Reagent A | Wako | Cat#: 999-34691 |
| HR Series NEFA-HR(2) Color Reagent B | Wako | Cat#: 991-34891 |
| HR Series NEFA-HR(2) Color Solvent A | Wako | Cat#: 995-34791 |
| HR Series NEFA-HR(2) Color Solvent B | Wako | Cat#: 993-35191 |
| Infinity™ Cholesterol Reagent | ThermoFisher | Cat#: TR13421 |
| Infinity™ Triglycerides Reagent | ThermoFisher | Cat#: TR22421 |
| iScript™ cDNA Synthesis Kit | BIO-RAD, | Cat#: 170-8891 |
| Mouse Adiponectin Elisa Kit | Millipore | Cat#: EZMADP-60K |
| Mouse Leptin ELISA Kit | Crystal Chem | Cat#: 90030 |
| Mouse Ultrasensitive Insulin ELISA | ALPCO | Cat#: 80-INSMSU-E01 |
| Oil red O stain kit | Abcam | Cat#: ab150678 |
| Pierce BCA Protein Assay kit | ThermoFisher | Cat#: 23225 |
| RIPA buffer | Sigma-Aldrich | Cat#: R0278 |
| RNeasy Mini Kit (Qiagen) | Qiagen | Cat#: 74106 |
| Serum iron/TIBC Kit | Pointe Scientific | Cat#: I750460 |
| Sybr Green Master Mix | Applied biosystems | Cat#: A25778 |
| TUNEL staining kit | ThermoFisher | Cat#: C10617 |
| Deposited data | ||
| RNA-seq | This paper | GEO: GSE144938 |
| 16S rRNA-seq | This paper | SRA NCBI: PRJNA718550 |
| Experimental models: Organisms/strains | ||
| Mouse: Tfrcfl/fl | The Jackson Laboratory | JAX028363; RRID:IMSR_JAX:028363 |
| Mouse: Adipoq-Cre | The Jackson Laboratory | JAX028020; RRID:IMSR_JAX:028020 |
| Mouse: Ucp1-Cre | The Jackson Laboratory | JAX024670; RRID:IMSR_JAX:024670 |
| Mouse: TRE-Cre | The Jackson Laboratory | JAX006234; RRID:IMSR_JAX:006234 |
| Mouse: Adipoq-rtTA | Sun et al., 2012 | N/A |
| Mouse: FAT-ATTAC | Pajvani et al., 2005 | N/A |
| Oligonucleotides | ||
| Primers for qPCR | See Table S5 | N/A |
| Recombinant DNA | ||
| pAAVK CAG-FLEX-eGFP | See Data S1 | N/A |
| pAAVK CAG-FLEX-Fpn1C326S | See Data S2 | N/A |
| pAAVK CAG-FLEX vector | See Data S3; Gift from Hongkui Zeng | Based on Addgene plasmid #51502 |
| pHelper | Cell Biolabs | Part No: 340202 |
| pRepCap Rec2 | Gift from Deborah Young, University of Auckland, New Zealand | N/A |
| pAAV-DJ | Cell Biolabs | Part No: VPK-420-DJ |
| Software and algorithms | ||
| Adobe photoshop | Adobe | https://www.adobe.com/products/photoshop.html; RRID: SCR_014199 |
| Endnote | Endnote | http://endnote.com; RRID: SCR_014001 |
| Excel | Microsoft | https://www.microsoft.com/enus/microsoft-365/excel; RRID:SCR_016137 |
| Image J | NIH | https://imagej.nih.gov/ij/; RRID: SCR_003070 |
| PowerPoint | Microsoft | https://www.microsoft.com/enus/microsoft-365/powerpoint |
| Prism 9 | GraphPad software | https://www.graphpad.com/; RRID: SCR_002798 |
| Word | Microsoft | https://www.microsoft.com/enus/microsoft-365/word |
| Other | ||
| 1200EX Transmission Electron Microscope | JEOL | N/A |
| 60% HFD Green pellet | Research diets | D12492i |
| 60% HFD paste | Bio-serv | S1850 |
| 6200 Isoperibol Calorimeter | Parr Instrument Company | N/A |
| a Nexera X2 UHPLC | Shimadzu Scientific Instruments | N/A |
| Agilent 2100 Bioanalyzer | Agilent | N/A |
| Amicon Ultra-15 centrifugal filter unit (3KD) | Millipore | UFC9003 |
| Amicon Ultra-2 centrifugal filter unit (3KD) | Millipore | UFC200324 |
| Beckman Coulter LS6500 multi-purpose scintillation counter | Beckman | N/A |
| BMDS IPTT reader | BMDS | Model: DAS-8007-IUS |
| Bruker Minispec mq10 | Brucker | Cat#: TMTA2892161 |
| DNA/RNA shield collection tube | ZYMO research | Cat#: R1102 |
| Doxycycline 60% HFD (600mg/kg diet) | Bio-serv | S5867 |
| Doxycycline chow diet (600 mg/kg diet) | Bio-serv | S4107 |
| Glucometers | Bayer Contour | N/A |
| Illumina Novaseq 6000 sequencing system | Illumina | N/A |
| Implantable electronic ID transponder IPTT-300 HTEC | BMDS | N/A |
| Keyence BZ-X710 Fluorescence Microscope | Keyence | N/A |
| LCMS-8050 | Shimadzu Scientific Instruments | N/A |
| Normal chow diet | TEKLAD | 2916 |
| Odyssey Infrared Imager | LI-COR Biosciences | N/A |
| PGA synthetic absorbable suture 5.0 | Henry Schein | 31995 |
| Rodent cold/thermo-Chamber | Powers Scientific | N/A |
| Scintillation vial | RPI | SKU: 121000CA |
| Seahorse XF Extracellualr Flux Analyzer | Agilent | N/A |
| Seahorse xFe24 islet capture fluxpak | Agilent | Cat#: 103518-100 |
| TSE calorimetric system | TSE system | N/A |