| Antibodies |
| anti β-Tubulin |
Cell signaling technology |
Cat. No. 2146; RRID: AB_2210545 |
| Phospho-HSL (Ser660) |
Cell signaling technology |
Cat. No. 4126; RRID: AB_490997 |
| Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody |
Cell signaling technology |
Cat. No. 9101; RRID: AB_331646 |
| goat anti-rabbit IgG-HRP |
SantaCruz |
Cat. No. sc-2004; RRID: AB_631746 |
| Biological Samples |
| 208 non-diabetic volunteers of American Indian ethnicity (63% male, mean age = 30 years, mean BMI 34 kg/m2). |
NIDDK |
N/A |
| Chemicals, Peptides, and Recombinant Proteins |
| SKF 96365 |
TOCRIS |
Cat. No. 1147 |
| KN-62 |
TOCRIS |
Cat. No. 1277 |
| AnCoA4 |
EMD Millipore |
Cat. No. 532999 |
| Uridine 5′-triphosphate trisodium salt hydrate |
SIGMA |
Cat. No. U6625 |
| Isoproterenol hydrochloride |
SIGMA |
Cat. No. I6504 |
| Ionomycin calcium salt |
SIGMA |
Cat. No. I3909 |
| Fluo-4, AM, cell permeant |
SIGMA |
Cat. No. F14201
|
| 3-Isobutyl-1-methylxanthine |
SIGMA |
Cat. No. I5879 |
| Dexamethasone |
SIGMA |
Cat. No. D1756 |
| Insulin from bovine pancreas |
SIGMA |
Cat. No. I6634 |
| Halt Phosphatase Inhibitor Cocktail |
Thermo Scientific |
Cat. No. 78420 |
| Halt Protease Inhibitor Cocktail, EDTA-free (100X) |
Thermo Scientific |
Cat. No. 78425 |
| Calcium-45 Radionuclide |
Perkin Elmer |
Cat. No. NEZ013001MC |
| Puromycin |
TAKARA |
Cat. No. 631306 |
| Polybrene Infection |
SIGMA |
Cat. No. TR-1003 |
| Collagen coating solutions |
Cell Applications, Inc. |
Cat. No. 125–50 |
| TLQP-21, human, mouse and R21A |
Cero et al.,2014 |
N/A |
| SB290157 |
SIGMA |
Cat. No. 1192 |
| C5a Receptor Agonist FKP - (D - Cha) - Cha - r |
Anaspec |
Cat. No. AS-65121 |
| W54011 |
Tocris |
Cat. No. 5455 |
| Primestar |
Takara/Fisher |
Cat. No. R045A |
| DpnI |
New England Biolabs |
Cat. No. R0176L |
| Blasticydin |
Invivogen |
Cat. No. ant-bl-10p |
| Hygromycin B |
KSE Scientific |
Cat. No. 15140–122 |
| BrightGlo |
Promega |
Cat. No. E2620 |
| DMEM |
VWR |
Cat. No. 45000–306 |
| Fetal Bovine Serum (FBS) |
VWR |
Cat. No. 97068–085 |
| 10xHBSS |
Invitrogen |
Cat. No. 14065–056 |
| Fatty acid free bovine serum albumin |
Sigma Aldrich |
Cat. No. A7030–10G |
| L-Ascorbic acid |
Sigma Aldrich |
Cat. No. A92902–25G |
| Critical Commercial Assays |
| Pierce BCA Protein Assay Kit |
Thermo Scientific |
Cat. No. 23225 |
| Free Glycerol Reagent |
SIGMA |
Cat. No. F6428 |
| RNeasy lipid tissue mini kit |
QIAGEN |
Cat. No. 74804 |
| RNase-Free DNase Set |
QIAGEN |
Cat. No. 79254 |
| iScript cDNA Synthesis Kit |
BIORAD |
Cat. No. 1708890 |
| iQ SYBR® Green Supermix |
BIORAD |
Cat. No. 1708880 |
| Amersham ECL Prime Western Blotting Detection Reagent |
GE health care |
RPN2236 |
| C3aR1 ShRNA plasmid |
SIGMA |
Cat. No. SHCLNG-NM_009779
|
| control ShRNA plasmid |
SIGMA |
SHC002 |
| Adipocyte differentiation tool kit |
ATCC |
ATCC® PCS-500–050 |
| RNeasy Micro Kit |
QIAGEN |
Cat. No. 74004 |
| Human Exon 1.0 ST Array microarray chip |
ThermoFisher |
Cat. No. 900651 |
| GeneChip Whole Transcript Sense Target Labeling Assay kit |
ThermoFisher |
Cat. No. 900652 |
| GeneChip Fluidics Station 450 |
ThermoFisher |
Cat. No. 00–0079 |
| GeneChip Scanner 3000 7G |
ThermoFisher |
Cat. No. 00–00213 |
| Experimental Models: Cell Lines |
| 3T3-L1 cells |
ATCC |
ATCC® CL-173 |
| 3T3-L1 cells |
Minnesota Obesity Center |
N/A |
| human mesenchymal stem cells |
ATCC |
ATCC® PCS-500–011 |
| HTLA |
A gift from Dr. Richard Axel, Columbia University |
N/A |
| Experimental Models: Organisms/Strains |
| 3 months old male WT mice on a mixed C57BL6/J and 129Sv (N = 6/group). |
Produced in house |
N/A |
| Recombinant DNA |
| C3aR1 Tango |
Kroeze et al., 2015 |
Addgene, Cat. No. 66231 |
| Software and Algorithms |
| Image lab |
BIORAD |
N/A |
| Molecular Modeling |
Small-Molecule Drug Discovery Suite 2018–4, Schrödinger, LLC, New York, NY, 2018 |
N/A |
| MD Simulation |
Desmond 2018–4, D. E. Shaw Research, New York, NY, 2018 |
N/A |
| MD Simulation Analysis |
Small-Molecule Drug Discovery Suite 2018–4, Schrödinger, LLC, New York, NY, 2018. |
N/A |
| Secondary structure analysis |
VMD 1.9.3, Theoretical and Computational Biophysics Group, NIH Center for Macromolecular Modeling and Bioinformatics, University of Illinois - Urbana-Champaign |
N/A |
| Evolutionary sequence alignment |
Constraint-based Multiple Alignment Tool (COBALT), National Center for Biotechnology Information |
N/A |
| Evolutionary analysis |
MrBayes, version 3.2 http://nbisweden.github.io/MrBayes/
|
N/A |
| Phylogenetic tree design |
Itol software. https://itol.embl.de
|
N/A |
| NIS-Elements |
Nikon |
N/A |
| Statistica 13.3 |
Tibco software Inc. |
N/A |
| Correlations were assessed using the statistical analysis system of the SAS institute |
Sas Institute, Cary, NC |
N/A |
| Prism |
GraphPad Software Inc. |
N/A |
