Key resources table
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| UCP1 | Abcam | Cat#AB10983 |
| total OXPHOS | Abcam | Cat#AB110413 |
| vinculin | Sigma | Cat#V9264 |
| beta-actin | Cell Signalling Technology | Cat#3700S |
| Anti-Mouse IgG HRP Conjugate | Promega | Cat#W4021 |
| Anti-Rabbit IgG HRP Conjugate | Promega | Cat#W4011 |
| CD4 (FITC; GK1.5) | Biolegend | Cat#100406 |
| CD19 (PerCP-Cy5.5; 6D5) | Biolegend | Cat#115534 |
| F4/80 (PE-CY7; BM8) | Biolegend | Cat#123114 |
| CD8a (APC/CY7; 53-6.7) | Biolegend | Cat#100713 |
| CD11b (EF450; M1/70) | eBioscience | Cat#48-0112-82 |
| CD3 (BV412; 17A2) | Biolegend | Cat#100227 |
| CD3 (BV605; 17A2) | Biolegend | Cat#100237 |
| CD11c (BV711; N418) | Biolegend | Cat#117349 |
| CD45 (BV785; 30-F11) | Biolegend | Cat#103149 |
| Fc-receptor-blocking antibody | eBioscience | Cat#14-0161-82 |
| CD4 (FITC; GK1.5) | Biolegend | Cat#100406 |
| CD19 (PerCP-Cy5.5; 6D5) | Biolegend | Cat#115534 |
| Bacterial and virus strains | ||
| Biological samples | ||
| Chemicals, peptides, and recombinant proteins | ||
| Cysteine-reactive phosphate tags (CPTs) | Synthesized in house | N/A |
| 10-plex TMT reagents | ThermoFisher Scientific | Cat#90406 |
| 16-plex TMT reagents | Thermo Fisher Scientific | Cat#A44520 |
| 2,4-Dinitrophenol | MilliporeSigma | Cat#D198501 |
| 3,3’,5-Triiodo-L-thyronine (T3) | Sigma-Aldrich | Cat#T2877 |
| 4X NuPAGE LDS sample buffer | Thermo Fisher Scientific | Cat#NP0007 |
| 90-day release 17-β estradiol pellets | Innovation research of America | Cat#NE-121 |
| Acetonitrile | Fisher Scientific | Cat# A9964 |
| Ammonium acetate | Sigma | Cat#372331-100G |
| Ammonium hydroxide | Sigma | Cat# 338818-100ML |
| Antimycin | Sigma | Cat#A8674 |
| Beta-mercaptoethanol | Sigma | Cat#M6250 |
| Bond-Breaker TCEP Solution, Neutral pH | ThermoFisher Scientific | Cat#77720 |
| Bovine Serum Albumin | Sigma | Cat#A9647 |
| Collagenase B | Sigma-Aldrich | Cat#COLLB-RO |
| collagenase type II | Worthington | Cat#LS0041771 |
| cOmplete Protease Inhibitor Cocktail | Sigma-Aldrich | Cat#CO-RO |
| D-glucose-13C6 | Cambridge Isotope | Cat#CLM-1396-PK |
| Dexamethasone | Sigma-Aldrich | Cat#D4902 |
| Dithiothreitol (DTT) | Sigma-Aldrich | Cat#DTT-RO |
| DMEM/F-12, GlutaMAX | Life Technologies | Cat#10565042 |
| DMEM/F-12+Glutamax | Invitrogen | Cat#10565-042 |
| EPPS | Sigma-Aldrich | Cat#E9502 |
| fatty acid free BSA | Sigma-Aldrich | Cat#A6003 |
| Fetal Bovine Serum | Gemini Bio-Products | Cat#100-106 |
| Fixation buffer | Biolegend | Cat#420801 |
| Formalin, Buffered, 10% | Fisher Scientific | Cat#SF100-4 |
| Glycocholate-d4 | Cambridge Isotope | Cat#DLM-2742 |
| GoTaq qPCR master mix | Promega | Cat#A6001 |
| Indomethacin | Sigma-Aldrich | Cat#I7378 |
| Inosine-15N4 | Cambridge Isotope | Cat#NLM-4264 |
| Insulin | Sigma-Aldrich | Cat#I5500 |
| Iodoacetamide | Sigma-Aldrich | Cat#I1149 |
| Isobutylmethylxanthine(IBMX) | Sigma-Aldrich | Cat#I7018 |
| Lambda phosphatase | Santa Cruz Biotechnology | Cat#sc-200312 |
| Lys-C | Wako Chemicals | Cat#125-05061 |
| Methanol | Fisher Scientific | Cat# A4124 |
| MitoQ | Mike Murphy | gifted |
| Norepinephrine | Sigma-Aldrich | Cat#A9512 |
| oligomycin | MilliporeSigma | Cat#80058-538 |
| Penicillin streptomycin | Fisher scientific | Cat#15-140-163 |
| Pierce ECL Western Blotting Substrate | Life Technologies | Cat#32106 |
| Rosiglitazone | Cayman | Cat#71740 |
| Rotenone | Sigma-Aldrich | Cat#R8875 |
| RPMI | Life Technologies | Cat#21870-076 |
| Thymine-d4 | Cambridge Isotope | Cat#DLM-1089 |
| Trizol | Life Technologies | Cat#15596026 |
| Trypan Blue | Sigma | Cat#T8154 |
| Trypsin | Promega | Cat#V5113 |
| Tween-20 | Sigma | Cat#P9416 |
| Zombie Aqua Live/dead stain | Biolegend | Cat#423101 |
| ddPCR Supermix | Bio-rad | Cat#186-3024 |
| DNeasy Blood & Tissue Kit | Qiagen | Cat #69504 |
| Critical commercial assays | ||
| High-Select Fe-NTA Phosphopeptide Enrichment Kit | ThermoFisher Scientific | Cat#A32992 |
| Pierce High pH Reversed-Phase Peptide Fractionation Kit | ThermoFisher Scientific | Cat#84868 |
| Micro BCA™ Protein Assay Kit | ThermoFisher Scientific | Cat#23235 |
| RNA Mini Kit | Invitrogen | Cat#12183025 |
| High-capacity cDNA reverse transcription kit | Fisher Scientific | Cat# 4368813 |
| PureLink RNA Mini Kit | Life Technologies | Cat#12183025 |
| Deposited data | ||
| Deposited proteomics data | PRIDE | PXD029270 |
| Experimental models: Cell lines | ||
| Experimental models: Organisms/strains | ||
| C253A mice | This paper | |
| Oligonucleotides | ||
| See Table S13 | ||
| Recombinant DNA | ||
| Software and algorithms | ||
| Xcalibur | ThermoFisher Scientific | Cat#OPTON-30965 |
| TraceFinder | ThermoFisher Scientific | Cat#OPTON-30688 |
| Adipocount | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5826178/) | |
| Adiposoft plugin | https://imagej.net/plugins/adiposoft | |
| AMIDE medical imaging software | http://amide.sourceforge.net/ | |
| Biorender | https://biorender.com/ | |
| Enrichr | Chen et al., 2013; Kuleshov et al., 2016 | https://amp.pharm.mssm.edu/Enrichr/ |
| FACSDiva software | BD Biosciences | |
| FLIR Tools software | https://support.flir.com/SwDownload/app/RssSWDownload.aspx?ID=1247 | FLIR Tools software |
| FlowJo v10 software | Tree Star | |
| ImageJ | NIH | https://imagej.nih.gov/ij/ |
| In-house mass spectrometry data analysis software | Huttlin et al., 2010 | N/A |
| NIS-Elements software | Nikon | |
| Prism 8 | GraphPad | https://www.graphpad.com/scientific-software/prism/ |
| SEQUEST | Eng et al., 1994 | https://doi.org/10.1016/1044-0305(94)80016-2 |
| BIORAD QuantaSoft software | Bio-rad | Cat#1864011 |
| Other | ||
| Q-Exactive HF-X mass spectrometer | ThermoFisher Scientific | Cat#0726042 |
| Quant Studio 6 Flex Real-Time PCR machine | Applied Biosciences | Cat# 44-856-99 |
| UltiMate 3000 HPLC | ThermoFisher Scientific | IQLAAAGABHFAPBMBFC |
| Seahorse XF24 Extracellular Flux Analyzer | Agilent | |
| Sep-Pak C18 Cartridges | Waters | Cat#WAT054955 |
| SunFire Prep C18 5 μm OBD column | Waters | Cat#186003969 |
| TCC-3000RS Thermostatted Column Compartment | ThermoFisher Scientific | Cat#5730.0000 |
| TissueLyser II | QIAGEN | Cat#85300 |
| Ultrasound machine | Philips | Affiniti70 |
| WPS-3000TBFC Biocompatible Well Plate Autosampler | ThermoFisher Scientific | Cat#5841.0020 |
| XF24 V7 cell culture microplates | Agilent | Cat#100777-004 |
| 2489 UV/Visible Detector | Waters | Cat#2489 |
| 2545 Binary Gradient Module | Waters | Cat#2545 |
| Count beads | Spherotech | Cat#ACFD-50-5 |
| EASY-nLC 1200 System | ThermoFisher Scientific | Cat#LC140 |
| EchoMRI (model ET-025) | Echo Medical Systems | |
| FAIMSPro | ThermoFisher Scientific | Cat#FMS02-10001 |
| FLIR thermal imaging camera (Model FLIR-T62101). | FLIR | model discontinued |
| Glucometer, ultra mini | One Touch | N/A |
| Heparin columns | Becton Dickinson | Cat# 365985 |
| High-fat diet | OpenSource Diets | Cat#D12492 |
| High-fat high-sucrose diet | OpenSource Diets | Cat#D09071702 |
| Agilent technologies 1200 series HPLC | Agilent | |
| LPG-3400RS Quaternary Pump w/Degasser | ThermoFisher Scientific | Cat#5040.0036 |
| LSRFortessa | BD Biosciences | |
| Luna 5 μm NH2 column | Phenomenex | Cat#00F-4378-B0 |
| MilliporeSigma™ Immobilon™-P PVDF Transfer Membranes | MilliporeSigma | Cat#IPVH00010 |
| Multi-Therm heat-shake | Sigma-Aldrich | Cat#Z755753 |
| Nanodrop 2000 | Thermo Fisher | Cat#ND-2000 |
| 1260 Infinity II LC System | Agilent | |
| NuPAGE™ 4 to 12%, Bis-Tris, 1.5 mm, Mini Protein Gel, 15-well | Thermo Fisher Scientific | Cat#NP0336BOX |
| Orbitrap Eclipse mass spectrometer | ThermoFisher Scientific | Cat#FSN04-10000 |
| Orbitrap Fusion Lumos Mass Spectrometer | ThermoFisher Scientific | Cat#IQLAAEGAAPFADBMBHQ |
| BIORAD C1000 Thermo Cycler | Bio-rad | Cat#1851197 |
| BIORAD ddPCR AutoDG | Bio-rad | Cat#1864101 |
| BIORAD QX200 Droplet Reader | Bio-rad | Cat#1864003 |