KEY RESOURCES TABLE.
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Rabbit monoclonal anti-ACSL1 | Cell Signaling Technology | Cat#9189S; RRID:AB_10891616 |
| Rabbit monoclonal anti-ACSL4 | Abcam | Cat#Ab155282; RRID:AB_2714020 |
| Rabbit monoclonal anti-beta-actin | Cell Signaling Technology | Cat#4970S; RRID:AB_2223172 |
| Rabbit monoclonal anti-Calreticulin | Abcam | Cat#Ab92516; RRID:AB_10562796 |
| Rabbit polyclonal anti-Cytochrome C | Cell Signaling Technology | Cat#4272; RRID:AB_2090454 |
| Rabbit monoclonal anti-GST | Cell Signaling Technology | Cat#2625S; RRID:AB_490796 |
| Rabbit monoclonal anti-HA | Cell Signaling Technology | Cat#3724; RRID:AB_1549585 |
| Rabbit monoclonal anti-IκBα | Cell Signaling Technology | Cat#4812S; RRID:AB_10694416 |
| Rabbit polyclonal anti-MTCO2 | Abcam | Cat#198286; RRID:AB_2861364 |
| Rabbit monoclonal anti-Myc | Cell Signaling Technology | Cat#2278; RRID:AB_490778 |
| Rabbit monoclonal anti-p65 | Cell Signaling Technology | Cat#8242T; RRID:AB_10859369 |
| Rabbit monoclonal anti-p-p65 (S536) | Cell Signaling Technology | Cat#3033S; RRID:AB_331284 |
| Rabbit polyclonal anti-TBK1 | Cell Signaling Technology | Cat#3031S; RRID:AB_2199749 |
| Rabbit monoclonal anti-pTBK1 (Ser172) | Cell Signaling Technology | Cat#5483S; RRID:AB_10693472 |
| Rabbit monoclonal anti-Tom20 | Cell Signaling Technology | Cat#42406; RRID:AB_2687663 |
| Anti-Flag M2 affinity gel | Sigma-Aldrich | Cat#A2220; RRID:AB_10063035 |
| Anti-Flag M2 magnetic beads | Sigma-Aldrich | Cat#M8823; RRID:AB_2637089 |
| Anti-HA magnetic bead | Thermo Scientific Pierce | Cat#88837; RRID:AB_2861399 |
| Anti-Myc magnetic bead | Thermo Scientific Pierce | Cat#88843; RRID:AB_2861398 |
| Bacterial and Virus Strains | ||
| Stellar Competent Cells | Clontech | Cat#636763 |
| GST-Full length TBK1 baculovirus | Shu et al., 2013 | N/A |
| GST-C-terminal truncated TBK1 baculovirus | Shu et al., 2013 | N/A |
| GST-C-terminal truncated TBK1 K38A baculovirus | Shu et al., 2013 | N/A |
| GST-C-terminal truncated TBK1 K38A baculovirus | Shu et al., 2013 | N/A |
| Chemicals, Peptides, and Recombinant Proteins | ||
| Amlexanox | Abcam | Cat#Ab142825 |
| Carnitine | Bachem Americas INC | Cat#F2700.0025 |
| Cayman 10576 | Cayman | Cat#10011249 |
| Coenzyme A sodium salt hydrate | Sigma-Aldrich | Cat#C3144 |
| Deuterium oxide (D2O) | Sigma-Aldrich | Cat#151882 |
| Fatty acid free BSA | Akron Biotechnology | Cat#AK8909–0100 |
| Liberase TM | Roche | Cat#5401127001 |
| Lipid standard, MG, DG, TG mix | Sigma-Aldrich | Cat#1787–1AMP |
| Octanoyl carnitine | Sigma-Aldrich | Cat#50892 |
| Palmitate-BSA | Agilent Technologies | Cat#102720–100 |
| 1-[C14]-palmitic acid | Perkin Elmer | Cat#NEC075H050UC |
| U-[C14]-palmitic acid | Perkin Elmer | Cat#NEC534050UC |
| Palmitoylcarnitine | Sigma-Aldrich | Cat#P1645 |
| Palmitoyl CoA | Sigma-Aldrich | Cat#P9716 |
| Percoll | Sigma-Aldrich | Cat#GE17089101 |
| Plasma Membrane Permeabilizer (PMP) | Agilent Technologies | Cat#102504–100 |
| Poloxamer 407 purified | Sigma-Aldrich | Cat#16758 |
| Protease and phosphatase inhibitor | Thermo Fisher Scientific | Cat#78444 |
| Proteinase K | Roche | Cat#03115836001 |
| Recombinant human TNFα | BioLegend | Cat#570106 |
| RNase A | Roche Diagnostics | Cat#10109169001 |
| Sodium palmitate | Sigma-Aldrich | Cat#P9767 |
| Triascin C | Enzo Life Sciences | Cat#BML-EI218–0100 |
| Critical Commercial Assays | ||
| BCA protein assay kits | Pierce | Cat#23227 |
| Cholesterol reagent | Thermo Fisher | Cat#TR13421 |
| CloneAmp Hifi DNA PCR premix | Takara | Cat#639298 |
| Free fatty acid uptake assay kit | Abcam | Cat#ab176768 |
| GST protein purification kit | Pierce | Cat#16107 |
| In-Fusion HD enzyme premix | Takara | Cat#639690 |
| Ketone body assay kit | Abnova | Cat#89028362 |
| Lipofectamine LTX | Life Technologies | Cat#A12621 |
| Lipofectamine p3000 | Life Technologies | Cat#L3000015 |
| Lipofectamine RNAiMAX | Life Technologies | Cat#13778500 |
| Profectine | AB vector | Cat#T10 |
| Maxima First Strand cDNA Synthesis kit | Thermo Scientific | Cat#K1642 |
| Power SYBR Green PCR Master Mix | Life Technologies | Cat#4367659 |
| Triglyceride reagent | Thermo Fisher | Cat#TR22421 |
| TRIzol reagent | Life Technologies | Cat#15596018 |
| Experimental Models: Cell Lines | ||
| HEK293 | ATCC | Cat#CRL-1573 RRID:CVCL_0045 |
| sf9 cells | Gibco | Cat#B82501 RRID:CVCL_0549 |
| Experimental Models: Organisms/Strains | ||
| Mouse: TBK1 f/f | Marchlik et al., 2010 | N/A |
| Mouse: Albumin Cre
B6.Cg-Speer6-ps1Tg(Alb-cre)21Mgn/J |
Jackson Laboratory | Cat#003574; RRID:IMSR_JAX:003574 |
| Mouse: C57BL/6J | Jackson Laboratory | Cat#000664; RRID:IMSR_JAX:000664 |
| Oligonucleotides | ||
| Mouse ACSL1 siRNA | Dharmacon | Cat#14081 |
| Negative control siRNA | Integrated DNA Technologies | Cat#51–01–14–04 |
| Primers for Q-PCR, see Table S1, S2 | ||
| Recombinant DNA | ||
| pcDNA-Flag-human TBK1 | Fitzgerald et al., 2003 | N/A |
| pcDNA-Flag-human TBK1 K38A | Fitzgerald et al., 2003 | N/A |
| pcDNA-Flag-human TBK1 S172A | This paper | N/A |
| 6myc-mTBK1 WT | Goncalves et al., 2011 | N/A |
| 6myc-mTBK1 ULD deleted | Goncalves et al., 2011 | N/A |
| 6myc-mTBK1 C-terminal deleted | Goncalves et al., 2011 | N/A |
| pCMV6 Entry mACSL1-myc/flag | Origene | Cat#MR210117 |
| pCMV6 Entry mACSL1-HA | This paper | N/A |
| Baculovirus genomic vector DNA | AB vector | Cat#A1 |
| pAcGHLTc-full length TBK1 | Shu et al., 2013 | N/A |
| pAcGHLTc-C-terminal truncated TBK1 | Shu et al., 2013 | N/A |
| pAcGHLTc-C-terminal truncated TBK1 K38A | Shu et al., 2013 | N/A |
| pAcGHLTc-C-terminal truncated TBK1 S172A | Shu et al., 2013 | N/A |
| Software and Algorithms | ||
| ImageJ | NIH | https://imagej.net/Welcome |
| GraphPad Prism 8.4.3 | GraphPad Software | https://www.graphpad.com/ |
| BioRender (for Graphical Abstract) | BioRender | https://www.biorender.com/ |
| Other | ||
| High Fat Diet (60% kcal% fat) | Research Diet, Inc. | Cat#D12492 |
| Methionine/Choline-deficient diet | Research Diet, Inc. | Cat#A06071302 |
| William’s medium E | Life Technologies | Cat#12551–032 |
| GlutaMax | Life Technologies | Cat#35050–061 |
| Spin columns | Abcam | Cat#ab93349 |
| TLC plate | EMD Millipore | Cat#1.05553.0001 |