| Antibodies |
| Anti-rabbit monoclonal phospho-GSK-3α (Ser21) (36E9) |
Cell Signaling |
9316 |
| Anti-rabbit monoclonal GSK-3α (D80E6) |
Cell Signaling |
4337 |
| Anti-rabbit monoclonal GSK-3α (D80D1) XP |
Cell Signaling |
4818 |
| Anti-rabbit monoclonal GSK-3α/β (D75D3) XP |
Cell Signaling |
5676 |
| Anti-rabbit polyclonal phospho-GSK-3α/β (Ser21/9) |
Cell Signaling |
9331 |
| Anti-rabbit monoclonal GSK-3β (27C10) |
Cell Signaling |
9315 |
| Anti-mouse monoclonal GSK-3α (H-12) |
Santa Cruz |
sc-5264 |
| Anti-rabbit polyclonal GSK-3β (H-76) |
Santa Cruz |
sc-9166 |
| Anti-rabbit polyclonal PPARα |
Cayman Chemical |
101710 |
| Anti-rabbit polyclonal PPARα |
Abcam |
ab24509 |
| Anti-rabbit polyclonal RXRα (D-20) |
Santa Cruz |
sc-553 |
| Anti-rabbit polyclonal RXRα/β/γ (ΔN197) |
Santa Cruz |
sc-774X |
| Anti-rabbit monoclonal β-Catenin (6B3) |
Cell Signaling |
9582 |
| Anti-rabbit polyclonal phospho-β-catenin (Ser33/37/T41) |
Cell Signaling |
9561 |
| Anti-rabbit polyclonal Akt |
Cell Signaling |
9272 |
| Anti-rabbit polyclonal phospho-Akt (Ser473) |
Cell Signaling |
9271 |
| Anti-rabbit polyclonal Histone H3 |
Cell Signaling |
9715 |
| Anti-rabbit monoclonal GAPDH (14C10) |
Cell Signaling |
2118 |
| Anti-rabbit monoclonal GFP (D5.1) XP |
Cell Signaling |
2956 |
| Anti-rabbit polyclonal GST |
Cell Signaling |
2622 |
| Anti-mouse monoclonal α-actinin (Sarcomeric) |
Sigma-Aldrich |
A7811 |
| Anti-mouse IgG, HRP-linked antibody |
Cell Signaling |
7076 |
| Anti-rabbit IgG, HRP-linked antibody |
Cell Signaling |
7074 |
| Anti-rabbit polyclonal Troponin T antibody |
Abcam |
45932 |
| Goat anti-mouse IgG secondary antibody, Alexa Fluor 488 |
Thermo Fisher Scientific |
A-11029 |
| Goat anti-rabbit IgG secondary antibody, Alexa Fluor 488 |
Thermo Fisher Scientific |
A-11034 |
| Goat anti-rabbit IgG secondary antibody, Alexa Fluor 594 |
Thermo Fisher Scientific |
A-11037 |
| Clean-Blot IP Detection Reagent (HRP) |
Thermo Fisher Scientific |
21230 |
| Anti-rabbit polyclonal phospho-specific PPARα (Ser280) |
GeneScript |
This paper |
| Bacterial and Virus Strains |
| Subcloning Efficiency DH5α Competent Cells |
Thermo Fisher Scientific |
18265017 |
| One Shot BL21(DE3) Chemically Competent E. coli
|
Thermo Fisher Scientific |
C600003 |
| BAC clone |
BACPAC |
RP24–545F24 |
| Biological Samples |
| Human heart samples |
Taipei Veterans General Hospital |
This paper |
| Chemicals, Peptides, and Recombinant Proteins |
| Anti-GFP mAb-Magnetic beads |
MBL |
D153–11 |
| Anti-FLAG M2 Affinity Gel |
Sigma-Aldrich |
A2220 |
| Protein A/G PLUS-Agarose |
Santa Cruz |
sc-2003 |
| Glutathione Sepharose 4B |
VWR/GE |
17–0756-01 |
| GSK3α protein, active |
Millipore |
14–492 |
| GSK3β protein, active |
Millipore |
14–306 |
| Recombinant GST-PPARa (full-length and truncated) |
This paper |
N/A |
| Recombinant GST-β-catenin |
This paper |
N/A |
| PPARα LBD (human recombinant) |
Cayman Chemical |
10009088 |
| Recombinant GSK-3α |
This paper |
N/A |
| IPTG |
Invitrogen |
15529–019 |
| FuGENE 6 Reagent |
VWR/Promega |
E2691 |
| LipofectamineTM 2000 Transfection Reagent |
Invitrogen |
11668–019 |
| Palmitic acid |
Sigma-Aldrich |
P0500 |
| Oleic acid |
Sigma-Aldrich |
O1008 |
| Linoleic acid |
Sigma-Aldrich |
L1376 |
| Bovine Serum Albumin |
Boston BioProducts |
P753 |
| Palmitic Acid, [9,10–3H(N)]-, 5mCi (185MBq) |
Perkin Elmer |
NET043005MC |
| Oleic Acid, [9,10–3H(N)]-, 1mCi |
Perkin Elmer |
NET289001MC |
| Etomoxir sodium salt hydrate |
Sigma-Aldrich |
E1905 |
| 2-Deoxy-D-glucose |
Sigma-Aldrich |
D8375 |
| L-Carnitine hydrochloride |
Sigma-Aldrich |
C0283 |
| Sodium Pyruvate |
Sigma-Aldrich |
P4562 |
| Fenofibrate |
Sigma-Aldrich |
F6020 |
| WY-14643 |
R&D Systems, Tocris |
1312 |
| Rodent Diet with 60% kcal% fat |
Research Diets |
D12492 |
| Rodent Diet with 10% kcal% fat |
Research Diets |
D12450B |
| High-fat diet containing 0.2% (wt/wt) fenofibrate |
Research Diets |
This paper |
| High-fat diet without Fenofibrate |
Research Diets |
This paper |
| Insulin solution human |
Sigma-Aldrich |
I9278 |
| 2,2,2-Tribromoethanol |
Sigma-Aldrich |
T48402 |
| Oil Red O solution in 0.5% in isopropanol |
Sigma-Aldrich |
O1391 |
| Phosphatase Inhibitor Cocktail 3 |
Sigma-Aldrich |
P0044 |
| Protease Inhibitor Cocktail |
Sigma-Aldrich |
P8340 |
| Maxima SYBR Green/ROX qPCR Master Mix (2x) |
Thermo/Fermentas |
K0223 |
| TRIzol Reagent (Ambion) |
Life Technologies |
15596–018 |
| Adenosine 5’-diphosphate sodium salt |
Sigma-Aldrich |
A2754 |
| Collagenase Type 2 CLS2 |
Worthington |
LS004177 |
| Percoll |
GE Healthcare |
17–0891-01 |
| Biotinylated Thrombin |
EMD Millipore |
69672–3 |
| Streptavidin Agarose |
EMD Millipore |
69203–3 |
| Critical Commercial Assays |
| XF24 Extracellular Flux Analyzer |
Seahorse Bioscience |
https://www.agilent.com/en/products/cell-analysis-(seahorse)/seahorse-analyzers |
| XF96 Extracellular Flux Analyzer |
Seahorse Bioscience |
https://www.agilent.com/en/products/cell-analysis-(seahorse)/seahorse-analyzers |
| BriteLite Plus |
Perkin Elmer |
6066761 |
| Luciferase assay system |
VWR/Promega |
E1501 |
| Duolink In Situ PLA probe anti-Mouse MINUS |
Sigma-Aldrich |
92004–0030 |
| Duolink In Situ PLA probe anti-Rabbit PLUS |
Sigma-Aldrich |
92002–0030 |
| TnT T7 Coupled Reticulocyte Lysate System |
VWR/Promega |
L4610 |
| QIAGEN HiSpeed Plasmid Maxi Kit |
QIAGEN |
12663 |
| QIAGEN HiSpeed Plasmid Midi Kit |
QIAGEN |
12643 |
| QIAprep Spin Miniprep Kit |
QIAGEN |
27104 |
| QIAquick PCR Purification Kit |
QIAGEN |
28106 |
| QIAquick Gel Extraction Kit |
QIAGEN |
28706 |
| Deposited Data |
| GSK-3α/β knock-in mice microarray data |
Matsuda, T. et al., 2008 |
GSE112160 |
| PPARα mutants RNaseq data |
This paper |
GSE112309 |
| Experimental Models: Cell Lines |
| H9C2 |
Oka, S. et al., 2011 |
N/A |
| HEK293 |
Maejima, Y. et al., 2013 |
N/A |
| Experimental Models: Organisms/Strains |
| Mouse: GSK-3α heterozygous floxed |
Gift from Dr. P.S. Klein (University of Pennsylvania) |
N/A |
| Mouse: GSK-3β floxed |
Gift from Dr. C.J. Phiel (Nationwide Children’s Hospital) |
He, F. et al., 2010. |
| Mouse: GSK-3α-Ser21Ala knock-in |
Gift from Dr. D.R. Alessi (University of Dundee) |
N/A |
| Mouse: GSK-3β-Ser9Ala knock-in |
Gift from Dr. D.R. Alessi (University of Dundee) |
N/A |
| Mouse: α-myosin heavy chain promoter-driven heterozygous Cre transgenic |
Gift from Dr. M.D. Schneider (Imperial College London) |
N/A |
| Mouse: α-myosin heavy chain promoter-driven FLAG-PPARα transgenic |
Gift from Dr. D. Kelly (University of Pennsylvania) |
N/A |
| Mouse: C57BL/6J |
Jackson Laboratory |
JAX 000664 |
| Mouse: ob/ob: B6.Cg-Lepob/J |
Jackson Laboratory |
JAX 000632 |
| Mouse: PPARα-Ser280Ala knock-in |
This paper |
N/A |
| Rat: Primary cultured neonatal ventricular cardiomyocytes |
Harlan |
1-day-Crl:(WI)BR-Wistar rats |
| Oligonucleotides |
| Oligonucleotides used in this study are provided in Table S5
|
This paper |
NCBI PRIMER-BLAST |
| Recombinant DNA |
| pDC316 shuttle vector |
Microbix |
N/A |
| pDC316-YFP shuttle vector |
Sadoshima lab |
Maejima, Y. et al., 2013 |
| pDC316-FLAG shuttle vector |
Sadoshima lab |
Maejima, Y. et al., 2013 |
| pDCSilencer |
Microbix |
N/A |
| pBHGloxΔE1.3 Cre |
Microbix |
N/A |
| pCold-GST |
Takara/Clontech |
3372 |
| CTNNB1 (untagged)-Human catenin, beta 1 |
Origene |
SC107921 |
| GSK3 alpha pMT2 |
Addgene |
15896 |
| pPPRE-tk-luciferase reporter |
Gift from Dr. Ronald Evans (Salk Institute) |
N/A |
| pCITE-GSK-3α |
This paper |
N/A |
| pL451 |
Gift from Dr. Takeda (Osaka University) |
N/A |
| pMCS-DTA |
Gift from Dr. Takeda (Osaka University) |
N/A |
| pMCS-Neo-PPARα-S280A-DTA |
This paper |
N/A |
| pCold-GST-PPARα-full length |
This paper |
N/A |
| pCold-GST-PPARα-truncate 1 |
This paper |
N/A |
| pCold-GST-PPARα-truncate 2 |
This paper |
N/A |
| pCold-GST-PPARα-truncate 3 |
This paper |
N/A |
| pCold-GST-PPARα-truncate 4 |
This paper |
N/A |
| pCold-GST-PPARα-truncate 5 |
This paper |
N/A |
| pDC316-YFP-PPARα-wild type |
This paper |
N/A |
| pDC316-YFP-PPARα-S280A |
This paper |
N/A |
| pDC316-YFP-PPARα-S280D |
This paper |
N/A |
| pDC316-YFP-PPARα-S280E |
This paper |
N/A |
| pDC316-YFP-PPARα-WT-H440A |
This paper |
N/A |
| pDC316-YFP-PPARα-WT-H440Q |
This paper |
N/A |
| pDC316-YFP-PPARα-WT-K448A |
This paper |
N/A |
| pDC316-YFP-PPARα-WT-K448N |
This paper |
N/A |
| pDC316-YFP-PPARα-WT-K449A |
This paper |
N/A |
| pDC316-YFP-PPARα-WT-K449N |
This paper |
N/A |
| pDC316-YFP-PPARα-WT-K448A/K449A |
This paper |
N/A |
| pDC316-YFP-PPARα-WT-K448N/K449N |
This paper |
N/A |
| pDC316-YFP-PPARα-WT-H440A/K448A/K449A |
This paper |
N/A |
| pDC316-YFP-PPARα-WT-H440Q/K448N/K449N |
This paper |
N/A |
| pDC316-YFP-PPARα-WT-H440A |
This paper |
N/A |
| pDC316-YFP-PPARα-S280D-H440Q |
This paper |
N/A |
| pDC316-YFP-PPARα-S280D-K448A |
This paper |
N/A |
| pDC316-YFP-PPARα-S280D-K448N |
This paper |
N/A |
| pDC316-YFP-PPARα-S280D-K449A |
This paper |
N/A |
| pDC316-YFP-PPARα-S280D-K449N |
This paper |
N/A |
| pDC316-YFP-PPARα-S280D-K448A/K449A |
This paper |
N/A |
| pDC316-YFP-PPARα-S280D-K448N/K449N |
This paper |
N/A |
| pDC316-YFP-PPARα-S280D-H440A/K448A/K449A |
This paper |
N/A |
| pDC316-YFP-PPARα-S280D-H440Q/K448N/K449N |
This paper |
N/A |
| pDC316-GSK-3α |
This paper |
N/A |
| shRNA-GSK-3α #1 |
This paper |
N/A |
| shRNA-GSK-3α #2 |
This paper |
N/A |
| shRNA-GSK-3α #3 |
This paper |
N/A |
| pBluescript II |
Maejima, Y. et al., 2013 |
N/A |
| AAV-DJ/8-CMV |
UPenn Vector Core |
N/A |
| AAV-DJ/8-CMV-PPARα-wild type |
This paper |
N/A |
| AAV-DJ/8-CMV-PPARα-S280D |
This paper |
N/A |
| Software and Algorithms |
| R version 3.3.1 |
The R Project |
https://www.r-project.org/ |
| CLC Genomics Workbench 8.5 version |
Qiagen |
http://www.clcbio.com/products/clc-genomics-workbench/ |
| GSEA ver 2.0 |
Broad institute of MIT and Harvard |
http://software.broadinstitute.org/gsea/ |
| IPA pathway analysis software |
Qiagen |
https://www.qiagenbioinformatics.com/products/ingenuity-pathway-analysis/ |
| MEME Suite ver 5.0.2 |
The MEME SUITE |
http://meme-suite.org/ |
| UCSC Genome Browser |
UCSC |
http://genome.ucsc.edu/ |
| ImageJ |
NIH |
https://imagej.nih.gov/ij/ |
| Prism version 6 |
GraphPad |
https://www.graphpad.com |
| Metaboanalyst ver 3.0 |
MetaboAnalyst |
http://www.metaboanalyst.ca/ |
| MultiExperiment Viewer ver4.9.0 |
MeV |
http://mev.tm4.org/#/welcome |
| Other |
|
|
|
