KEY RESOURCES TABLE
REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
Rabbit monoclonal p-AKT (Ser473) (D9E) | Cell Signaling | 4060; RRID:AB_2315049 |
Rabbit monoclonal pan AKT (C67E7) | Cell Signaling | 4691; RRID:AB_915783 |
Rabbit polyclonal adiponectin | GeneTex | GTX112777; RRID:AB_2885323 |
Mouse monoclonal Beta actin (AC-15) | Sigma-Aldrich | A5441; RRID:AB_476744 |
Rabbit polyclonal HSP90 | GeneTex | GTX110089; RRID:AB_1950529 |
Armenian hamster monoclonal CD11c (N418) | ThermoFisher | 12-0114-82; RRID:AB_465552 |
Rat monoclonal CD206 (MR5D3) | Bio-Rad | MCA2235F; RRID:AB_324594 |
Rat monoclonal CD45 (30-F11) | ThermoFisher | 17-0451-82; RRID:AB_469392 |
Rat monoclonal F4/80 (BM8) | ThermoFisher | 45-4801-82; RRID:AB_914345 |
Rabbit polyclonal p-HSL (Ser563) | Cell Signaling | 4139; RRID:AB_2135495 |
Rabbit polyclonal total HSL | Cell Signaling | 4107; RRID:AB_2296900 |
Rat monoclonal Galectin-3 (MAC2) (M3/38) | eBioscience | 13-5301-82; RRID:AB_837113 |
Rat monoclonal Galectin-3 (MAC2) (M3/38) | Novus Biologicals | NBP1-43313; RRID:AB_10006681 |
Rabbit monoclonal GAPDH (14C10) | Cell Signaling | 2118; RRID:AB_561053 |
Guinea pig Perilipin 1 | Progen | GP-29; RRID:AB_2892611 |
Rabbit polyclonal Tyrosine hydroxylase | Pel-Freez Biologicals | P40101-150; RRID:AB_2617184 |
Rabbit monoclonal pSTAT3 (Tyr705) (D3A7) | Cell Signaling | 9145; RRID:AB_2491009 |
Mouse monoclonal CD68 (KP1) | Invitrogen | MA5-13324; RRID:AB_10987212 |
Donkey anti-mouse IgG Alexa Fluor 488 | Jackson ImmunoResearch Labs | 715-545-151; RRID:AB_2341099 |
Goat anti-rabbit IgG Alexa Fluor 488 | ThermoFisher | A-11034; RRID:AB_2576217 |
Donkey anti-rat Alexa Fluor 647 | Jackson ImmunoResearch Labs | 712-606-153; RRID:AB_2340696 |
Donkey anti-guinea pig IgG Alexa Fluor 647 | Jackson ImmunoResearch Labs | 706-605-148; RRID:AB_2340476 |
IRDye 800CW donkey anti-rabbit IgG | Li-Cor | 925-32213; RRID AB_2715510 |
IRDye 680RD donkey anti-mouse IgG | Li-Cor | 925-68072; RRID AB_2814912 |
Bacterial and virus strains | ||
Biological samples | ||
Human subcutaneous preadipocytes | Zen-Bio Inc. | #SL0065 |
Chemicals, peptides, and recombinant proteins | ||
Auranofin | Sigma-Aldrich | A6733 |
Tamoxifen | MP Biomedicals | 156738 |
T-PER Tissue Protein Extraction Reagent | ThermoFisher | 78510 |
Halt Protease and Phosphatase Inhibitor Cocktail | ThermoFisher | 78440 |
4′,6-diamidino-2-phenylindole (DAPI) | Sigma-Aldrich | D8417 |
MitoTracker Red CMX-ROS | ThermoFisher | M7512 |
LipidTOX | Life Technologies | H34475 |
DAPI Fluoromount-G | VWR | 0100-20 |
IRDye 680RD Streptavidin | LI-COR | 926-68079 |
Tissue-Tek OCT compound | Sakura Finetek | 4583 |
Formaldehyde | ThermoFisher | 28908 |
Collagenase type I | Worthington Biochemical Corp | LS004194 |
Collagenase type I | Roche | 17100-017 |
Dispase II | Sigma-Aldrich | D4693 |
Isobutylmethylxanthine (IBMX) | Sigma-Aldrich | 13347 |
Rosiglitazone | Cayman Chemical Co | 71740 |
Dexamethasone | Tocris Biosciences | 1126 |
Insulin | Sigma-Aldrich | I5500 |
3,3′,5-Triiodo-L-thyronine (T3) | Sigma-Aldrich | T-074 |
DMSO | Sigma-Aldrich | D2660 |
Polyethylene glycol 400 | Affymetrix | 19957 |
Ethanol absolute 200 proof | Fisher Scientific | BP2818 |
10% Neutral buffered formalin | Fisher Chemical | SF100-4 |
DMEM/F-12, GlutaMAX | ThermoFisher | 10565-018 |
Fetal Bovine Serum | VWR | 97068-085 |
Penicillin-Streptomycin | Gibco | 15140-122 |
Sodium pyruvate | Gibco | 11360-070 |
L-glutamine | Gibco | 25030-081 |
Seahorse XF assay medium modified DMEM | Agilent | 102365-100 |
Isoproterenol | Cayman Chemical | 15592 |
2-deoxy-D-glucose, [1-14C] | PerkinElmer | NEC720A050UC |
Glucose, D-[2-3H] | PerkinElmer | NET238C001MC |
Critical commercial assays | ||
Rat/Mouse Insulin ELISA | Millipore | EZRMI-13K |
Mouse Leptin ELISA kit | Crystal Chem | 90030 |
Adiponectin Mouse ELISA Kit | ThermoFisher | KMP0041 |
DPPIV (DPP4/CD26) Mouse ELISA Kit | ThermoFisher | EMDPP4 |
Proteome Profiler Mouse Adipokine Array Kit | R&D Systems | ARY013 |
Triglycerides Reagent | ThermoFisher | TR22421 |
Glycerol Assay kit | Sigma-Aldrich | MAK117 |
Serum/Plasma Fatty Acid Detection Kit | Zen-Bio Inc. | sfa-1 |
Deposited data | ||
RNA-seq data | This paper | GEO: GSE202935 |
Experimental models: Cell lines | ||
Experimental models: Organisms/strains | ||
Mouse: C57/Bl6J | BCM Center for Comparative Medicine | N/A |
Mouse: FVB | BCM Center for Comparative Medicine | N/A |
Mouse: Ubc-CreERT2;LepR-lp/lp | Cox et al. 2016 | N/A |
Mouse: B6.Cg-Lepob/J | Jackson Laboratory | 000632; RRID:IMSR_JAX:000632 |
Beta-less (bAR1, bAR2, bAR3 knockout mice) | Bachmann et al. 2002; Xu et al. 2014 | N/A |
miR-30a −/− | This paper | N/A |
Oligonucleotides | ||
Primers for qPCR, see Table S2 | This paper | N/A |
sgRNA sequence: miR-30a upstream ACTCCCGCAGAGCACTTCTCAGG | This paper | N/A |
sgRNA sequence: miR-30a downstream TCAAGGAGTAATTATCTTGTTGG | This paper | N/A |
Primer: miR-30a forward GCATCGAGGCTTTGCAGTTT | This paper | N/A |
Primer: miR-30a reverse TGCACAGGAAGAACACTTCTGT | This paper | N/A |
Recombinant DNA | ||
Software and algorithms | ||
Fiji | Schindelin et. al. 2012 | https://imagej.net/software/fiji/ |
ImageJ with Adiposoft plugin | CIMA, University of Navarra | https://imagej.net/plugins/adiposoft |
Prism 9 | GraphPad | https://www.graphpad.com/ |
CalR | Palmer et al. 2017 | https://calrapp.org/ |
STAR | Dobin et al., 2013 | https://github.com/alexdobin/STAR |
DESeq2 | Love et al., 2014 | https://github.com/mikelove/DESeq2 |
GSEA | Subramanian et al., 2005 | https://www.gsea-msigdb.org/gsea/index.jsp |
GenePix Pro 7.0 Microarray Acquisition & Analysis Software | Molecular Devices | https://www.moleculardevices.com |
Other | ||
4-12% Bis-Tris NuPage gels | Life Technologies | NP0321Box |
Immobilon-P Transfer Membranes | Millipore | IPVH00010 |
Direct-zol RNA MiniPrep kit | Zymo Research | R2051 |
qScript | QuantBio | 95048-100 |
Sso-Advanced Universal Probes Supermix | Bio-Rad | 175284 |
TaqMan Advanced miRNA cDNA Synthesis | ThermoFisher | A28007 |
TaqMan Advanced miRNA Assays | ThermoFisher | A25576 |
TaqMan Gene Expression Assays (FAM) | ThermoFisher | 4331182 |
NEBNext Ultra II DNA library prep kit | New England Biolabs | E7645S |
Seahorse XF24 Islet Capture FluxPak | Agilent | 103518-100 |
Seahorse XF24 FluxPak | Agilent | 102342-100 |
Seahorse XF24 V7-PS cell culture microplates | Agilent | 100777-004 |
Seahorse XF Cell Mito Stress Test Kit | Agilent | 103010-100 |
60% high fat diet | Bio-Serv | S3282 |
Precision Count Beads | BioLegend | 424902 |