KEY RESOURCES TABLE
REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
Rabbit polyclonal anti-ATP citrate lyase | Proteintech | Cat#15421-1-AP; RRID:AB_2223741 |
Mouse monoclonal anti-β-actin | Cell Signaling Technology | Cat#3700S; RRID:AB_2242334 |
Rabbit monoclonal anti-AceCS1 | Cell Signaling Technology | Cat#3658T |
Rabbit monoclonal anti-pyruvate dehydrogenase | Cell Signaling Technology | Cat#3205T; RRID:AB_2162926 |
Rabbit polyclonal anti-aKGDH | Cell Signaling Technology | Cat#26865S |
Rabbit monoclonal anti-citrate synthase | Cell Signaling Technology | Cat#14309S |
Rabbit monoclonal anti-MPC1 | Cell Signaling Technology | Cat#14462S |
Rabbit polyclonal anti-Lamin A/C | Cell Signaling Technology | Cat#2032T |
Rabbit polyclonal anti-Histone H3 | Cell Signaling Technology | Cat#9715S; RRID:AB_331563 |
Rabbit monoclonal anti-acetyl-histone K27 | Cell Signaling Technology | Cat#8173T |
Rabbit monoclonal anti-acetyl-histone K9 | Cell Signaling Technology | Cat#9649T;RRID:AB_823528 |
Rabbit polyclonal anti-acetyl-histone H3 | Millipore | Cat#06-599; RRID:AB_2115283 |
Goat anti-mouse IgG (H&L) | Rockland Immunochemicals | Cat#610-1102; RRID:AB_219646 |
Donkey anti-rabbit IgG (H&L) peroxidase conjugated | Rockland Immunochemicals | Cat#611-7302; RRID:AB_219747 |
Chemicals, Peptides, and Recombinant Proteins | ||
RPMI 1640 | Cellgro | Cat#11875-093 |
RPMI 1640, without glucose | ThermoFisher Scientific | Cat#11879020 |
RPMI 1640, without glucose, without sodium bicarbonate | Sigma-Aldrich | Cat#R1383-10X1L |
RPMI 1640, without thiamine | This paper | N/A |
Opti-MEM | ThermoFisher Scientific | Cat#11058021 |
Fetal Bovine Serum (FBS) | Hyclone | Cat#14-486 |
Dialyzed FBS | ThermoFisher Scientific | Cat#H00000406-P01 |
[13C6]-D-glucose | Cambridge Isotope Laboratories | Cat#CLM-1396-1 |
3,3,3-[2H3]-sodium pyruvate | Cambridge Isotope Laboratories | Cat#DLM-6068-0.5 |
[13C3]-sodium pyruvate | Cambridge Isotope Laboratories | Cat#CLM-2440-0.5 |
3,3,3-[2H3]-sodium lactate | Cambridge Isotope Laboratories | Cat#DLM-9071-0.1 |
[2H3]-acetyl-carnitine hydrochloride | Santa Cruz Biotechnology | Cat#sc-480972 |
[2H3]-acetyl-CoA | Laboratory of Hening Lin, Cornell University | (Sadhukhan et al., 2016) |
18O2 | Sigma-Aldrich | Cat#490474-1L |
2,2,2-[2H3]-sodium acetate | Sigma-Aldrich | Cat#176079-5G |
[2H4]-acetaldehyde | Sigma-Aldrich | Cat#176567-1G |
[2H2]-H2O | Sigma-Aldrich | Cat#151882-10G |
[18O2]-H2O2 (2-3 % solution) | Sigma-Aldrich | Cat#609978 |
Coenzyme A sodium salt hydrate | Sigma-Aldrich | Cat#C3144-25MG |
β-Nicotinamide adenine dinucleotide sodium salt (NAD+) | Sigma-Aldrich | Cat#N0632-1G |
Glutathione (reduced) | Sigma-Aldrich | Cat#G4251-1G |
SAHA | Sigma-Aldrich | Cat#SML0061 |
Ammonium tetrathiomolybdate (TTM) | Sigma-Aldrich | Cat#323446 |
CPI-613 | Sigma-Aldrich | Cat#SML0404 |
UK5099 | Sigma-Aldrich | Cat#PZ0160 |
Copper (II) sulfate | Sigma-Aldrich | Cat#451657 |
2-hydrazinoquinoline | Sigma-Aldrich | Cat#CDS000062 |
Triphenylphosphine | Sigma-Aldrich | Cat#T84409 |
2,2’-dipyridyl disulfide | Sigma-Aldrich | Cat#143049 |
L-Kynurenine | Sigma-Aldrich | Cat#K8625 |
L-Methionine sulfoxide | Sigma-Aldrich | Cat#M1126 |
Pyruvate dehydrogenase (PDH) from porcine heart | Sigma-Aldrich | Cat#P7032-10UN |
alpha-Ketoglutarate dehydrogenase (aKGDH) from porcine heart | Sigma-Aldrich | Cat#K1502-20UN |
Catalase from human erythrocytes | Sigma-Aldrich | Cat#C3556 |
Thiamine pyrophosphate chloride | Alfa Aesar | Cat#J61483 |
Ac-CoA Synthase Inhibitor | Millipore | Cat#533756 |
MTT powder | ThermoFisher Scientific | Cat#M6494 |
Quick Start™ Bradford 1x Dye Reagent | Bio-Rad | Cat#5000205 |
Lipofectamine™ RNAiMAX Transfection Reagent | ThermoFisher Scientific | Cat#13778030 |
Ponceau S solution, 0.1% | G-Biosciences | Cat#786-576 |
cOmplete™, EDTA-free Protease Inhibitor Cocktail | Sigma-Aldrich | Cat#11873580001 |
IGEPAL CA-630 | Sigma-Aldrich | Cat#I8896 |
Acetonitrile, Optima LC/MS | Fisher Scientific | Cat# A955 |
Water, Optima LC/MS | Fisher Scientific | Cat# W6 |
Methanol, Optima LC/MS | Fisher Scientific | Cat# A456 |
Ammonium hydroxide, Optima LC/MS | Fisher Scientific | Cat#A470 |
Formic acid, Optima LC/MS | Fisher Scientific | Cat#A117 |
Acetic Acid, Glacial | Fisher Scientific | Cat#A38 |
Ammonium hydroxide, Optima LC/MS | Fisher Scientific | Cat#A470 |
Critical Commercial Assays | ||
Mitochondria Isolation Kit for Cultured Cells | ThermoFisher Scientific | Cat#89874 |
Western Lightning Plus-ECL, Enhanced Chemiluminescence Substrate | Perkin Elmer | Cat#NEL103001EA |
Deposited Data | ||
Histone proteomics data | This paper; Mendeley Data | http://dx.doi.org/10.17632/39rzr8xzsv.1 |
Metabolomics data | This paper; Mendeley Data | http://dx.doi.org/10.17632/39rzr8xzsv.1 |
NMR data | This paper; Mendeley Data | http://dx.doi.org/10.17632/39rzr8xzsv.1 |
Experimental Models: Cell Lines | ||
Human: HCT116 cells | ATCC | CCL-247 |
Human: HepG2 cells | ATCC | HB-8065 |
Human: SKOV cells | ATCC | HTB-77 |
Human: BT474 cells | ATCC | HTB-20 |
Human: HeyA8 cells | Laboratory of Ernst Lengyel, University of Chicago | (Kenny et al., 2015) |
Mouse: Sarcoma cells | This paper | N/A |
Mouse: MEF (ACLY WT and KO) cells | Laboratory of Kathryn Wellen, University of Pennsylvania | (Zhao et al., 2016) |
Experimental Models: Organisms/Strains | ||
Mouse: C57BL/6J | The Jackson Laboratory | Stock No. 000664 |
Mouse: Pax7CreER-T2 | Laboratory of Chen-Ming Fan, Carnegic Institution for Science | N/A |
Mouse: p53fl/fl | Laboratory of Anton Berns, The Netherlands Cancer Institute | N/A |
Mouse: LSL-NrasG12D | The Jackson Laboratory | Stock No. 008304 |
Oligonucleotides | ||
siRNA pools for human ACLY | Dharmacon | Cat#L-004915-00 |
siRNA pools for human ACSS2 | Dharmacon | Cat#L-010396-00 |
siRNA pools for mouse ACSS2 | Dharmacon | Cat# L-065412-01 |
Negative control siRNA | Dharmacon | Cat#D-001810-10-20 |
Recombinant DNA | ||
LentiCRISPR v2 expressing an sgRNA targeting PDH (5’-AGAACAACCGCTATGGCATG-3’) | (Sanjana et al., 2014) | Addgene Plasmid#52961 |
Software and Algorithms | ||
Sieve 2.0 | ThermoFisher Scientific | https://portal.thermo-brims.com/index.php/component/thermosoftwares/thermosoftware/67?Itemid=121 |
Xcalibur 4.0 | ThermoFisher Scientific | https://www.thermofisher.com/order/catalog/product/OPTON-30487 |
MestReNova Lite SE | Mestrelab Research | http://mestrelab.com/news/new-mnova-lite-10-special-edition/ |
GraphPad Prism 6 | GraphPad Software | https://www.graphpad.com/scientific-software/prism/ |
ChemDraw 15 | PerkinElmer | http://www.cambridgesoft.com/land/ChemDraw_Q22014.aspx |
Microsoft Office Excel | Microsoft | https://www.microsoft.com/en-us/p/excel-2016/cfq7ttc0k5f3?activetab=pivot%3aoverviewtab |
Other | ||
Mini vacuum desiccator | VWR | Cat#47750-242 |