KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Goat anti-Mouse polyclonal IgG HRP-linked antibody | Santa Cruz | Cat$sc-2005; RRID:AB_631736 |
| Goat anti-rabbit IgG (H+L) Antibody, Alexa Fluor 488 | ThermoFisher Scientific | Cat#A32731; RRID:AB_2633280 |
| Goat anti-rabbit IgG (H+L) Antibody, Alexa Fluor 568 | Molecular Probes | Cat#A-11004; RRID:AB_2534072 |
| Goat anti-Rabbit IgG HRP-linked antibody | Cell Signaling | Cat#7074S, RRID:AB_2099233 |
| Mouse monoclonal anti-DDIT3 (L63F7) | Cell Signaling | Cat#2895; RRID:AB_2089254 |
| Mouse monoclonal anti-INSRbeta (C-19) | Santa Cruz | Cat#sc-711; RRID:AB_631835 |
| Mouse monoclonal anti-SQSTM1/P62 | Abnova | Cat#H00008878-M03; RRID:AB_1112274 |
| Rabbit monoclonal anti-ADGRE1/F4/80 (D2S9R) | Cell Signaling | CAT#70076; RRID:AB_2799771 |
| Rabbit monoclonal anti-ATG7 | Abgent | Cat#AP1813a; RRID:AB_636747 |
| Rabbit monoclonal anti-EIF2AK3(D11A8) | Cell Signaling | Cat#5683, RRID:AB_10841299 |
| Rabbit monoclonal anti-FOXO1 (C29H4) | Cell Signaling | Cat#76764; RRID:AB_2799887 |
| Rabbit monoclonal anti-HSPA5(C50B12) | Cell Signaling | Cat#3177S; RRID:AB_2119845 |
| Rabbit monoclonal anti-Phospho-AKT1 (Ser473) (D9E) | Cell Signaling | Cat#4060; RRID:AB_2315049 |
| Rabbit monoclonal anti-phospho-EIF2AK3 (Thr980) | Cell Signaling | Cat#3179S; RRID:AB_2095853 |
| Rabbit monoclonal anti-TUBA (11H10) | Cell Signaling | Cat#2125S; RRID:AB_2619646 |
| Rabbit polyclonal anti-ACTB(H-300) | Santa Cruz | Cat#sc-10731; RRID:AB_2223515 |
| Rabbit polyclonal anti-ADH5 | Abcam | Cat#91385; RRID:AB_2049142 |
| Rabbit polyclonal anti-AKT1 2/3 (H-136) | Santa Cruz | Cat#sc-8312; RRID:AB_671714 |
| Rabbit polyclonal anti-ATG5 | Novus | Cat#NB110-53818; RRID:AB_828587 |
| Rabbit polyclonal anti-Histone H3K27me3 Active Motif | ThermoFisher Scientific | Cat#39156; RRID:AB_2867260 |
| Rabbit polyclonal anti-HSF1 | Cell Signaling | Cat#4356S; RRID:AB_2120258 |
| Rabbit polyclonal anti-MAP1LC3B | Novus | Cat#NB600-1384; RRID:AB_669581 |
| Rabbit polyclonal anti-phospho-FOXO1 (Ser256) | Cell Signaling | Cat#9461; RRID:AB_329831 |
| Rabbit polyclonal anti-UCP1 | Abcam | Cat#ab10983; RRID:AB_2241462 |
| TMT Monoclonal Antibody (25D5) | ThermoFisher Scientific | Cat#90075; RRID:AB_10854708 |
| Bacterial and virus strains | ||
| Adeno-Adh5 | Dr. Ling Yang; Qian et al., 2018 | N/A |
| Adeno-GFP | Yang et al., 2015 | N/A |
| Chemicals, peptides, and recombinant proteins | ||
| 1400W | Sigma | Cat#214358-33-5 |
| 3-Isobutyl-1 -methylxanthine (IBMX) | Sigma | Cat#I5879 |
| 3,3′-Diaminobenzidine (DAB) Liquid Substrate System tetrahydrochloride | Sigma | Cat#D7304-1SET |
| 5-Aza-2′-Deoxycytidine (5aza) | Sigma | Cat#189825 |
| Adenosine 5′-diphosphate bis(cyclohexylammonium) salt (ADP) | Sigma | Cat#A4386 |
| CL316, 243 | Sigma | Cat#C5976 |
| DAPI (4′,6-diamidino-2-phenylindole) | ThermoFisher Scientific | Cat#D1306 |
| Dexamethasone | Sigma | Cat#D4902 |
| Guanosine 5′-diphosphate sodium salt (GDP) | Sigma | Cat#G7127 |
| HSF1A | Millipore | Cat#1196723-93-9 |
| Humulin R U-500 | Lily | Cat#0002-8501-01 |
| Indomethacin | Sigma | Cat#4025268 |
| Iodoacetyl Tandem Mass TagTM (iodoTMTTM) Reagents | ThermoFisher Scienticic | Cat#90102 |
| Isoproterenol | Sigma | Cat#54750-10-6 |
| KRIBB11 | Sigma | Cat#385570 |
| Liberase | Roche | Cat#5401119001 |
| Lipofectamine 2000 | ThermoFisher Scientific | Cat#L30000015 |
| Malate | Sigma | Cat#W237418 |
| N6022 | Sigma | Cat#1208315-24-5 |
| NDC 0409-6648-02 DEXTROSE | Hospira | Cat#0409-6648-02 |
| Recombinant human UCP1 | MyBiosource | Cat#MBS963514 |
| Rosiglitazone | Sigma | Cat#557366-M |
| S-Nitroso-N-acetyl-DL-penicillamine (SNAP) | Sigma | Cat#N3398 |
| Sodium pyruvate | Sigma | Cat#P5280 |
| Succinate | Sigma | Cat#W239607 |
| Critical commercial assays | ||
| Free Fatty Acid Fluorometric Assay Kit | Cayman Chemical | Cat#700310 |
| Luciferase Assay | Promega | Cat#E1500 |
| NBEBuilder HiFi DNA assembly system | NEB | Cat# E5520S |
| Nitrate/Nitrite Colorimetric Assay Kit | Cayman Chemical | Cat#780001 |
| Pierce S-Nitrosylation Western Blot Kit | ThermoFisher Scientific | Cat#90105 |
| Renilla Luciferase Assay | Promega | Cat#E2810 |
| VECTASTAIN® ABC-HRP Kit, Peroxidase (Standard) PK-4000 | Vector Laboratories | Cat#PK-4000 |
| Experimental models: Cell lines | ||
| Human: A41hBAT-SVF | ATCC | Cat#CRL-3385 |
| Human: HEK293A | ATCC | Cat#CRL-1573 |
| Mouse: Brown Adipocytes | Dr. Ling Yang | N/A |
| Mouse: Stromal Vascular Fraction | Dr. Ling Yang | N/A |
| Experimental models: Organisms/strains | ||
| Mouse: Adh5BKO | This paper | N/A |
| Mouse; Adh5fl | Dr. Limin Liu; Wei et al.; 2011 | N/A |
| Mouse: C57BL/6J | The Jackson Laboratory | JAX: Cat#000664 |
| Mouse: FVB-Tg(UCP1-cre)1Evdr/J | The Jackson Laboratory | JAX: Cat#024670 |
| Oligonucleotides. See Table S1. | ||
| Recombinant DNA | ||
| Plasmid:C305AUCP1 | This paper | N/A |
| Plasmid:GFP | This paper | N/A |
| Plasmid:pcDNA3.1-IDH2-WT | Addgene | Cat#V790-20 |
| Plasmid:WTUCP1 | This paper | N/A |
| Software and algorithms | ||
| GraphPad Prism 9.20 | GraphPad Software, LLC. | https://www.graphpad.com/scientific-software/prism/ |
| Image Studio Lite | LI-COR Biosciences | https://www.licor.com/bio/image-studio-lite/ |
| ImageJ | Schneider et al., 2012 | https://imagej.nih.gov/ij/ |
| Other | ||
| 60%kCal high-fat diet | Research Diets | Cat#D12492 |
| Oil Red O staining | Sigma | Cat#O0625 |
| Opti-MEM | ThermoFisher Scientific | Cat#31985-062 |
| Teklad global soy protein-free extruded 2920X | Teklad Global Diet | N/A |