KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| mTOR | Cell Signaling Technology | Cat# 2972; RRID: AB_330978 |
| RICTOR | Cell Signaling Technology | Cat# 2140; RRID: AB_561245 |
| PPARγ (Western blot) | Cell Signaling Technology | Cat# 2443; RRID: AB_823598 |
| Perilipin1 (IF staining) | Cell Signaling Technology | Cat# 9349; RRID: AB_10829911 |
| C/EBP0α | Santa Cruz Biotechnology | Cat# sc-365318; RRID: AB_10846948 |
| α-Tubulin | Cell Signaling Technology | Cat# 2125; RRID: AB_2619646 |
| Phospho-S473-AKT | Cell Signaling Technology | Cat# 4058; RRID: AB_331168 |
| Phospho-T450-AKT | Cell Signaling Technology | Cat# 9267; RRID: AB_823676 |
| Phospho-T308-AKT | Cell Signaling Technology | Cat# 4056; RRID: AB_331163 |
| Phospho-S473-AKT1 | Cell Signaling Technology | Cat# 9018; RRID: AB_2629283 |
| Phospho-S474-AKT2 | Cell Signaling Technology | Cat# 8599; RRID: AB_2630347 |
| AKT1 | Cell Signaling Technology | Cat# 2938; RRID: AB_915788 |
| AKT2 | Cell Signaling Technology | Cat# 3063; RRID: AB_2225186 |
| AKT | Cell Signaling Technology | Cat# 9272; RRID: AB_329827 |
| Phospho-T24-FoxO1 | Cell Signaling Technology | Cat# 9464; RRID: AB_329842 |
| FoxO1 | Cell Signaling Technology | Cat# 2880; RRID: AB_2106495 |
| Phospho-S9-GSK3β | Cell Signaling Technology | Cat# 9322; RRID: AB_2115196 |
| GSK3bβ | Cell Signaling Technology | Cat# 12456; RRID: AB_2636978 |
| Phospho-T246-PRAS40 | Cell Signaling Technology | Cat# 2997; RRID: AB_2258110 |
| PRAS40 | Cell Signaling Technology | Cat# 2691; RRID: AB_2225033 |
| Phospho-T642-AS160 | Cell Signaling Technology | Cat# 8881; RRID: AB_2651042 |
| AS160 | EMD Millipore | Cat# 07–741; RRID: AB_492639 |
| Phospho-T389-S6K1 | Cell Signaling Technology | Cat# 9205; RRID: AB_330944 |
| S6K1 | Santa Cruz Biotechnology | Cat# sc8418 RRID: AB_628094 |
| Insulin Receptor(IR)β | Cell Signaling Technology | Cat# 3025; RRID: AB_2280448 |
| ACC | Cell Signaling Technology | Cat# 3676; RRID: AB_2219397 |
| ACLY | Cell Signaling Technology | Cat# 4332; RRID: AB_2223744 |
| FASN | Cell Signaling Technology | Cat# 3180; RRID: AB_2100796 |
| ChREBP | Novus | Cat# NB400–135; RRID: AB_10002435 |
| SREBP | EMD Millipore | Cat# 04–469, RRID:AB_612072 |
| HA | Cell Signaling Technology | Cat# 2367; RRID: AB_10691311 |
| Histone H3 | Cell Signaling Technology | Cat# 4499; RRID: AB_10544537 |
| Acetyl-Histone H3 (lys9) | Cell Signaling Technology | Cat# 9649; RRID: AB_823528 |
| PPARg | Santa Cruz Biotechnology | Cat# sc-7196; RRID: AB_654710 |
| CD36 | NOVUS | Cat#: NB400–144SS; RRID: AB_920879 |
| SCD1 | Abclonal | Cat# A16429 |
| FABP4 | Cell Signaling Technology | Cat# 2120; RRID: AB_2102466 |
| Goat anti-Rabbit IgG (H+L) CrossAdsorbed Secondary Antibody, Alexa Fluor 568 | Thermo Fisher Scientific | Cat# A-11011; RRID: AB_143157 |
| PE-Cy7-conjugated anti-CD31 | eBioscience, Thermo Fisher Scientific | Cat# 25-0311-82; RRID: AB_2716949 |
| PE-Cy7-conjugated anti-CD45 | eBioscience, Thermo Fisher Scientific | Cat# 25-0451-82; RRID: AB_2734986 |
| A700-conjugated anti-CD29 | BioLegend | Cat# 102218; RRID: AB_493711 |
| A647-conjugated anti-CD34 | BioLegend | Cat# 119314; RRID: AB_604089 |
| LybA/E-conjugated anti-Sca1 | eBioscience, Thermo Fisher Scientific | Cat# 45-0242-82; RRID: AB_1210701 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| 4-hydroxy Tamoxifen (4-OHT) | Toronto research chemicals | H954729 |
| Rosiglitazone | Cayman Chemical | 71740 |
| Oil Red O | Sigma-Aldrich | O0625 |
| Human insulin, regular | Novo Nordisk | #183302 |
| Insulin | Sigma-Aldrich | I2643 |
| 3-isobutyl-1-methylxanthine (IBMX) | Sigma-Aldrich | I5879 |
| Dexamethasone | Sigma-Aldrich | D1756 |
| Osmium tetroxide 2% aqueous solution | Polysciences Inc | #23311 |
| HCS LipidTOX Deep Red Neutral Lipid Stain | Invitrogen | H34477 |
| Trichostatin A (TSA) | Sigma-Aldrich | T1952 |
| BODIPYFLC16 | Invitrogen | D3821 |
| Propidium iodide | Invitrogen | P3566 |
| Critical Commercial Assays | ||
| Dual-Luciferase Reporter assay system | Promega | E1910 |
| Free Glycerol Reagent | Sigma-Aldrich | F6428 |
| NEBNext® Ultra Directional RNA Library Prep Kit for Illumina® | New England Biolabs | E7760 |
| Seahorse XF Glycolysis stress test kit | Agilent Technologies | Cat# 103020-100 |
| Seahorse XF Palmitate Oxidation stress test kit and FAO substrate | Agilent Technologies | Cat# 103693-100 Cat# 102720-100 |
| Deposited Data | ||
| Raw and analyzed data | This study | GSE146470 |
| Experimental Models: Cell Lines | ||
| Immortalized subcutaneous white preadipocytes | (Tang et al., 2016) | N/A |
| Primary subcutaneous white preadipocytes | This study | N/A |
| Experimental Models: Organisms/Strains | ||
| Mouse: Prx1-Cre | Jackson Labs | 005584 |
| Mouse: Adiponectin-Cre | Jackson Labs | 028020 |
| Mouse: Rictorfloxed | Jackson Labs | 020649 |
| Mouse: UBC-creERT2 | Jackson Labs | 007001 |
| Oligonucleotides | ||
| Mouse primers | IDT | See Table S4 |
| Recombinant DNA | ||
| pMSCV-Puro | (Akama-Garren et al., 2016) | RRID: Addgene_68469 |
| pMSCV-ChREBPα-Puro | (Witte etal., 2015) | N/A |
| pMSCV-ChREBPβ-Puro | (Witte etal., 2015) | N/A |
| pMSCV-HA-PPARγ2-Hygro | This study | N/A |
| pMSCV-Myc-ACLY-Puro | (Calejman et al., 2020) | N/A |
| pMSCV-Myc-ACLY-S455D-Puro | (Calejman et al., 2020) | N/A |
| pMSCV-Myc-ACLY-S455E-Puro | This study | N/A |
| pMSCV-Myc-ACLY-S455A-Puro | (Calejman et al., 2020) | N/A |
| PPRE X3-TK-luc | (Kim etal., 1998) | RRID: Addgene_1015 |
| pMSCV-HA-AKT1-Hygro | (Calejman et al., 2020) | N/A |
| pMSCV-HA-AKT1-S473D-Hygro | (Calejman et al., 2020) | N/A |
| pMSCV-HA-AKT1-S473A-Hygro | (Calejman et al., 2020) | N/A |
| pMSCV-HA-AKT2-Hygro | (Calejman et al., 2020) | N/A |
| pMSCV-HA-AKT2-S474D-Hygro | (Calejman et al., 2020) | N/A |
| pSV Sport PPAR gamma 1 | Addgene | RRID: Addgene_8886 |
| pSV Sport PPAR gamma 2 | Addgene | RRID: Addgene_8862 |
| Software and Algorithms | ||
| ImageJ (Fiji) | (Schindelin et al., 2012) | https://imagej.nih.gov/ij/ |
| Adiposoft Fiji plugin) | (Galarraga et al., 2012) | N/A |
| star_2.5.3a | (Dobin et al., 2013) | N/A |
| Ensembl annotation GRCm38.94 | (Zerbino et al., 2018) | N/A |
| featureCounts_1.5.2 | (Liao, Smyth, and Shi 2014) | N/A |
| DESeq2_1.20.0 | (Love, Huber, and Anders 2014) | N/A |
| Other | ||
| ChIP-Atlas | (Okietal. 2018) | N/A |
| DAVID Bioinformatics Resources 6.8 | (Huang, Sherman, and Lempicki 2009a; 2009b) | https://david.ncifcrf.gov/home.jsp |
| Cytation™ 5 Image reader | Biotek | N/A |
| LSRII A-5 laser flow cytometer | BD Biosciences | N/A |
| Seahorse XFe96 Analyzer | Agilent | N/A |