KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Hamster monoclonal anti-mouse CD3 | BioXCell, eBioscience | BE0001-1 |
| Hamster monoclonal anti-mouse CD28 | BioXCell, eBioscience | BE00015-1 |
| Hamster monoclonal anti-mouse IFN-gamma | BioXCell | BE0055 |
| Rat monoclonal anti-mouse IL-4 | BioXCell | BE0045 |
| Rat monoclonal anti-mouse CD25 | eBioscience | Clone PC61.5 |
| Rat monoclonal anti-mouse CD4 | eBioscience | Clone GK1.5 |
| Rat monoclonal anti-mouse CD62L | eBioscience | Clone MEL-14 |
| Rat monoclonal anti-mouse Foxp3 | eBioscience | Clone FJK-16s |
| Rat monoclonal anti-mouse IL-17A | eBioscience | Clone eBio17B7 |
| Rat monoclonal anti-mouse TNF-alpha | eBioscience | Clone MP6-XT22 |
| Rat monoclonal anti-mouse IFN-gamma | eBioscience | Clone XMG1.2 |
| Rat monoclonal anti-mouse CD8 | eBioscience | Clone 53-6.7 |
| Mouse monoclonal anti-mouse CD45.2 | eBioscience | Clone 104 |
| Rat monoclonal anti-mouse CD127 | BioLegend | Clone A7R34 |
| Hamster monoclonal anti-mouse KLRG1 | eBioscience | Clone 2F1 |
| Rat monoclonal anti-mouse CD44 | eBioscience | Clone IM7 |
| Mouse monoclonal anti-human/mouse CPT1A | Abcam | Clone 8F6AE9 |
| Mouse monoclonal anti-human/mouse Foxp3 | eBioscience | eBio7979 |
| AMPKα (23A3) Rabbit mAb | Cell Signaling | #2603 |
| Phospho-AMPKα (Thr172) Rabbit mAb | Cell Signaling | #2535 |
| Anti-rabbit IgG, HRP-linked Antibody | Cell Signaling | #7074 |
| Anti-mouse IgG, HRP-linked Antibody | Cell Signaling | #7076 |
| Rabbit polyclonal anti-human/mouse CPT1B | Abcam | # ab104662 |
| Rabbit polyclonal anti-human/mouse CPT1C | Proteintech | # 12969-1-AP |
| Rabbit polyclonal anti-human/mouse beta-actin | Cell Signaling | # 4967 |
| H-2Kb/OVA257 Tetramer | Baylor College of Medicine | |
| Bacterial and Virus Strains | ||
| Recombinant Listeria monocytogenes (rLM-OVA) | Derived from Lm strain 10403S | |
| recombinant attenuated Listeria monocytogenes (Lm) deleted for actA and expressing OVA (attLmOVA) | ||
| recombinant virulent LmOVA (virLmOVA) | ||
| Chemicals, Peptides, and Recombinant Proteins | ||
| RPMI 1640 GlutaMAX | Thermo Fisher | 61870-010 |
| RPMI Medium 1640 (no glucose) | Thermo Fisher | 11879-020 |
| IMDM GlutaMAX | Thermo Fisher | 3198-022 |
| Heat-inactivated FCS | Biochrom | S 0115 - Lot# 1289W |
| Penicillin-streptomycin | Millipore | A2213 |
| 2-mercaptoethanol | Gibco | #21985-023 |
| recombinant human TGF-beta | Peprotech | 100-21 |
| recombinant murine IL-6 | Peprotech | 216-16 |
| recombinant murine IL-1beta | Peprotech | 211-11B |
| recombinant human IL-2 | Roche | C-149447 |
| recombinant murine IL-12 | Peprotech | 210-12 |
| recombinant murine IL-7 | Peprotech | 217-17 |
| Etomoxir sodium salt hydrate | Sigma Aldrich | E1905 |
| Phorbol 12-myristate 13-acetate (PMA) | Sigma-Aldrich | P-8139 |
| Ionomycin | Sigma-Aldrich | 10634 |
| Tetramethylrhodamine, Ethyl Ester, Perchlorate (TMRE) | Invitrogen | T669 |
| Pierce RIPA Buffer | Thermo Scientific | |
| Oligomycin | Sigma Aldrich | 75351 |
| FCCP | Cayman | 15218 |
| Rotenone | Cayman | 13955 |
| Antimycin A | Sigma Aldrich | A8674 |
| XF Base Medium Minimal DMEM without Phenol Red | Agilent | 103335-100 |
| XF Plasma Membrane Permeabilizer | Agilent | |
| L-carnitine | Sigma Aldrich | C0158 |
| Glucose ≥99,5 % D(+) | Roth | HN06.1 |
| L-Glutamine | Biochrom AG | M11-004 |
| Sodium Pyruvate | Gibco | #11360-070 |
| Oligo(dT)18-22 Primers | Invitrogen | |
| IQ SYBR Green Supermix | Bio-Rad | #170-8882 |
| Palmitic acid | Sigma Aldrich | P0500 |
| Palmitic acid (U-13C16) | Cambridge Isotope Laboratories | CLM-409-0.5 |
| Poly-D-lysine hydrobromide | Sigma Aldrich | P6407 |
| RNAse OUT Ribonuclease inhibitor (recombinant | Invitrogen | #10777-019 |
| Albumin fraction V fatty acid free | Roth | 0052.1 |
| Recombinant murine IL-15 | Peprotech | #3210-15 |
| Fixable Viability Dye eFluor 506 | eBioscience | #65-0866-14 |
| Fixable Viability Dye eFluor 780 | eBioscience | #65-0865-14 |
| Cell Proliferation Dye eFluor 450 | eBioscience | #65-0842-85 |
| Sodium Palmitate | Sigma Aldrich | P9767 |
| (U-13C16) Sodium Palmitate | Sigma Aldrich | 700258 |
| Sodium Butyrate | Sigma Aldrich | B5887 |
| BD GolgiStop | BD Biosciences | #51-2092KZ |
| Foxp3/transcription factor staining buffer set | eBioscience | #00-5523-00 |
| OVA(257-264) SIINFEKL peptide | Bio-Synthesis Inc. | custom |
| LLO(91–99) GYKDGNEYI peptide | Bio-Synthesis Inc. | custom |
| Critical Commercial Assays | ||
| EasySep Mouse T Cell Isolation Kit | StemCell technologies | #19851 |
| EasySep Mouse CD4 T cell isolation kit | StemCell technologies | #19852 |
| EasySep Mouse CD8 T cell isolation kit | StemCell technologies | #19853 |
| Dynabeads Untouched Mouse CD4 Cells Kit | Invitrogen | 11415D |
| Dynabeads Untouched Mouse CD8 Cells Kit | Invitrogen | 11417D |
| Cell Trace Violet Cell Proliferation Kit | Invitrogen | C34571 |
| eBioscience™ Annexin V Apoptosis Detection Kit eFluor™ 450 | eBioscience | 888006-72 |
| LIVE/DEAD™ Fixable Aqua Dead Cell Stain Kit, for 405 nm excitation | Invitrogen | L34957 |
| Seahorse XF Palmitate-BSA FAO Substrate | Agilent Technologies | #102720-100 |
| Seahorse XFe96 FluxPak | Agilent technologies | #102416-100 |
| Qiagen RNAeasy Kit | Qiagen | #74106 |
| SuperScript III Reverse Transcriptase | Invitrogen | #18080-044 |
| High Capacity cDNA Reverse Transcription Kit | Applied Biosystems | #4368814 |
| SensiFAST SYBR Lo-Rox Mix | Bioloine | #BIO-94050 |
| Experimental Models: Organisms/Strains | ||
| Cpt1afl/fl mice: C57BL/6NTac.(129S6/SvEvTac;C57BL/6NCr)F1-Cpt1atm1 Pec | Schoors et al., Nature 520(7546):192-7 (2015) | |
| ACC2ko mice: B6.Cg-Acacb(tm1Sjw) | Baylor College of Medicine, Houston, texas, USA (Abu-Elheiga et al, 2001, Science, 291, 2613–2616) | |
| LCAD mice: B6.129S6-Acadltm1Uab | Dr. Philip Wood, Burnham Institute for Medical Research(Kurtz et al, PNAS, 1998, 22;95(26):15592-7) | |
| VLCADD mice: C57Bl/6-acadvltm1AS | Dr. Arnold Strauss Cincinnati Children’s Hospital Medical Center (Exil, et al 2003, Circulation Research. 2003;93:448–455) | |
| Oligonucleotides | ||
|
Cpt1a primer Fwd: 5’ CTCCGCCTGAGCCATGAAG 3’ Rev: 5’ CACCAGTGATGATGCCATTCT 3’ |
PrimerBank | ID 27804309a1 |
|
Cpt1b primer Fwd: 5’ GCACACCAGGCAGTAGCTTT 3’ Rev:5’ CAGGAGTTGATTCCAGACAGGTA 3’ |
PrimerBank | ID 6753512a1 |
|
Cpt1c primer Fwd: 5’ GATTTTCTGTACGTCACCCCC 3’ Rev: 5’ AGGTGGTCTTTTTCCACCCC 3’ |
This paper | |
|
Actb primer Fwd: 5’ TGT TAC CAA CTG GGA CGA CA 3’ Rev: 5’ GGG GTG TTG AAG GTC TCA AA 3’ |
This paper | |
|
Foxp3 primer Fwd: 5’ AGAAGCTGGGAGCTATGCAG 3’ Rev: 5’ GCTACGATGCAGCAAGAGC 3’ |
This paper | |
|
Cpt1a primer - IL-2/IL-15 cultures Fwd: 5’ CCAACGGGCTCATCTTCTAATC 3’ Rev: 5’ TGGTACAAAGCACCCATTAC 3’ |
Invitrogen | custom |
|
Cpt1b primer - IL-2/IL-15 cultures Fwd: 5’ ATTCTGTGCGGCCCTTATT 3’ Rev: 5’ TGACTTGAGCACCAGGTATTT 3’ |
Invitrogen | custom |
|
Cpt1c primer - IL-2/IL-15 cultures Fwd: 5’ GTTTCCTCTGGAGGTGGATTT 3’ Rev: 5’ GCTGGAGATATGGAAGGTGATT 3’ |
Invitrogen | custom |
|
OGDH primer - IL-2/IL-15 cultures Fwd: 5’ AATGAGGAGAAGCGGACCTT 3’ Rev: 5’ TCCACTCCATTTGCACTTGA 3’ |
Invitrogen | custom |
|
Prkaa1 primer - Fwd: 5’ CAGAGATCGGGATCCATCAG 3’ Rev: 5’ CCTGGTCTTGGAGCTACGTC 3’ |
This paper | |
| PR8 NP primer 5’ CAGCCTAATCAGACCAAATG 3’ 5’ TACCTGGTTCTCAGTTCAAG 3’ |
Tarasenko et al 2017 Cell metabolism | |
| Software and Algorithms | ||
| FlowJo 9.9.5 | FlowJo LLC | www.flowjo.com |
| GraphPad Prism V6 or V7 | GraphPad Software | www.graphpad.com |
| Other | ||