Key Resources Table
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| UCP1 | Abcam | ab23841 |
| F4/80 | Novus biologicals | NB600-404 |
| TNF alpha | Abcam | ab6671 |
| CD36 | Abcam | ab133625 |
| PGC1 alpha | Abcam | ab54481 |
| CPT1B | Abcam | ab134988 |
| PPAR alpha | Abcam | ab24509 |
| UCP3 | Abcam | ab3477 |
| Insulin Receptor | Abcam | ab137747 |
| IRS1 | Abcam | ab52167 |
| β-Tubulin | Sigma | T8328 |
| AMPK alpha 2 | Abcam | ab3760 |
| AMPK alpha 2 (phospho S491) | Abcam | ab109402 |
| PPAR gamma | Abcam | ab45036 |
| Acetyl-CoA Carboxylase | Cell Signaling | #3662 |
| Phospho-Acetyl-CoA Carboxylase | Cell Signaling | #3661 |
| PRDM16 | Abcam | ab202344 |
| SIRT1 | Abcam | ab12193 |
| IL-1β (3A6) | Cell Signaling | #12242 |
| SREBP1 | Novus biologicals | NB100-2215 |
| SREBP1 (Phospho-S439) | Biorbyt | orb315711 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| Taqman PCR Master Mix, no AmpErase UNG | Applied Biosystems | Cat# 4324018 |
| PALMITIC ACID, [1-14C]-, 50 μCi |
Perkin Elmer | NEC075H050UC |
| Dexamethasone | Sigma | D4902 |
| Indomethacin | Sigma | I7378 |
| 3-Isobutyl-1-methylxanthine | Sigma | I7018 |
| Insulin | Sigma | I2643 |
| Oil Red O | Sigma | O0625 |
| Dibutyryl cyclic-AMP | Sigma | D0260 |
| AICAR | Sigma | A9978 |
| Critical Commercial Assays | ||
| Dual-Luciferase® Reporter | Promega | E1910 |
| Assay System | ||
| Transcriptor reverse transcriptase | Roche | Cat# 03531287001 |
| RNeasy Plus Mini Kit | QIAGEN | Cat# 74134 |
| Pierce BCA Protein Assay Kit | Thermo Scientific | Cat# 23225 |
| Seahorse Bioassay | Agilent Technologies | Cat# 102416 |
| QuikChange™ Site-Directed Mutagenesis Kit | Agilent Technologies | Cat# 200518 |
| Experimental Models: Cell Lines and Primary cells | ||
| HepG2 | ATCC | ATCC® HB-8065™ |
| 3T3L1 | Dr. Kai Ge | ATCC® CL-173™ |
| C2C12 | ATCC | ATCC® CRL-1772™ |
| Brown preadipocytes | Dr. Kai Ge | Wang et al., 2010 |
| Primary human white preadipocyte | Dr. Susan K. Fried | |
| Primary preadipocytes | This study | |
| Recombinant DNA | ||
| PPARg | GeneCopoeia | MmiT054223 |
| PPARa | GeneCopoeia | MmiT029952 |
| PGC1a | GeneCopoeia | MmiT055331 |
| PRDM16 | GeneCopoeia | MmiT035098 |
| Oligonucleotides | ||
| PPARa-3’ UTR muta | cagagtggtttgtttgtttgtttttgttttaaaaatcgtatcattaagatatagctatgaagagcattctttttcattc gaatgaaaaagaatgctcttcatagctatatcttaatgatacgatttttaaaacaaaaacaaacaaacaaaccactctg | |
| PGC1a-3’UTR muta | Atattttgatgggctacccatcgcttctgcataagatacacaagcttcactaattttagatgca tgcatctaaaattagtgaagcttgtgtatcttatgcagaagcgatgggtagcccatcaaaatat | |
| PPARg-3’UTR muta | Ctaaaactttttgtttttaaatgctttttcatattaaatttcttaggtgtcagatttttttccctcaaaataat attattttgagggaaaaaaatctgacacctaagaaatttaatatgaaaaagcatttaaaaacaaaaagttttag | |
| Prdm16-3’UTR muta | gactgtccctagccctgagcgatgataggacaagacattcttgaattctc gagaattcaagaatgtcttgtcctatcatcgctcagggctagggacagtc | |