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. Author manuscript; available in PMC: 2020 Jul 10.
Published in final edited form as: Cell Rep. 2019 Apr 30;27(5):1376–1386.e6. doi: 10.1016/j.celrep.2019.04.005

KEY RESOURCES TABLE.

REAGENT or RESOURCE SOURCE IDENTIFIER
Antibodies
Anti-Lipoic acid Millipore 437695-100UL; RRID:AB_212120
Anti-LIPT1 Sigma-Aldrich AV48784; RRID:AB_1852908
Anti-DLAT Cell Signaling #12362; RRID:AB_2797893
Anti-DLAT Santa Cruz sc-271534; RRID:AB_10649809
Anti-DLST Cell Signaling #5556; RRID:AB_10695157
Anti-DBT GeneTex GTX113412; RRID:AB_2036741
Anti-p-PDHA1(Ser293) Millipore ABS204; RRID:AB_11213668
Anti-PDHA1 Cell Signaling #3205; RRID:AB_2162926
Anti-ACTB Sigma-Aldrich A3854; RRID:AB_262011
Anti-GAPDH Cell Signaling #8884; RRID:AB_11129865
Biological Samples
Patients’ blood samples This paper N/A
Patient’s skin biopsies This paper N/A
Chemicals, Peptides, and Recombinant Proteins
Puromycin ThermoFisher A1113803
Etomoxir sodium salt hydrate Sigma-Aldrich E1905
α-Ketoglutarate Sigma-Aldrich K1128
Citrate Sigma-Aldrich 251275
Fumarate Sigma-Aldrich F19353
Malate Sigma-Aldrich 240176
Succinate Sigma-Aldrich S3674
L-Aspartate Sigma-Aldrich A7219
Adenosine Sigma-Aldrich A9251
Guanosine Sigma-Aldrich G6264
Uridine Sigma-Aldrich U3003
[U-13C5]Glutamine Cambridge Isotope Laboratories CLM-1822
[U-13C6]Glucose Cambridge Isotope Laboratories CLM-1396
[1-14C]Pyruvate PerkinElmer NEC255050UC
L-[U-14C]Valine PerkinElmer NEC291EU050UC
3H2O PerkinElmer 6004052
Critical Commercial Assays
Kit Lenti-Smart INT Invivogen ItIns-10
Kit mammalian mitochondria isolation BioVision K288
Kit PDH activity colorimetric assay BioVision K679
Deproteinizing Sample Preparation Kit BioVision K808
Kit PicoProbeAcetyl-CoA Fluorometric Assay BioVision K317
Kit Q5 Site-Directed Mutagenesis Kit NEB E0554S
Kit CellTiter-Glo luminescent cell viability assay Promega G7572
Deposited Data
Human LIPT1 mutation N44S ClinVar SCV000588895
Human LIPT1 mutation S292X ClinVar SCV000588895
Experimental Models: Cell Lines
Patient-derived fibroblast and ctrl-4917 control fibroblast This paper N/A
Primary normal human neonatal fibroblasts ATCC PCS-201-010
Primary normal human adult fibroblasts ATCC PCS-201-012
Detroit 551 ATCC CCL-110
BJ ATCC CRL-2522
H460 ATCC HTB-177
Experimental Models: Organisms/Strains
B6C3F1/J mice The Jackson Laboratory 100010
Lipt1N44S/+ mice This paper N/A
Oligonucleotides
Genotyping primer LIPT1-N44S forward: 5′-AAAGCCAAAAGCAACTGCAC-3′ This paper N/A
Genotyping primer LIPT1-N44S reverse: 5′-GCCAAGGATTTTGATGCCTA-3′ This paper N/A
Genotyping primer LIPT1-S292X forward: 5′-AAACCCAACGGATGAGACAC-3′ This paper N/A
Genotyping primer LIPT1-S292X reverse: 5′-TGTTTAGTTTGTGGTCTTGTGGA-3′ This paper N/A
Cloning primer LIPT1 forward: 5′-CCGGCGATCGCATGCTGATCCCATTTTC –3′ This paper N/A
Cloning primer LIPT1 reverse: 5′-CGGACGCGTCATTATTCCCTTAATTTTTTC-3′ This paper N/A
Cloning primer LIPT1-S292X reverse: 5′-CGGACGCGTCTGTTCATATAACACATGAAAGG-3′ This paper N/A
Mutagenesis primer LIPT1-N44S forward: 5′-GTCTATCAAAGTCTGGCTGTG-3′ This paper N/A
Mutagenesis primer LIPT1-N44S reverse: 5′-ATCATTGGAAATTGACTGTAAAATG-3′ This paper N/A
Genotyping primer Lipt1-N44S forward: 5′-AAGAAGGTCTGGACCCTTTGA-3′ This paper N/A
Genotyping primer Lipt1-N44S reverse: 5′-GCCAGTTTGATTCCTTCCTG-3′ This paper N/A
sgRNA target sequence of Lipt1 for CRISPR knock-in of N44S: 5′-CTTCTAGATGTATGTGGTCGTGG-3′ This paper N/A
HR template single-strand oligo for CRISPR knock-in of N44S: 5′-CTTTGTCAGCACAAGGTCCCAGCAGCTGGCTTTAAAAGCCCACCCACACATGGGCTCATTTTACAGTCGATCTCCAATGATGTATATGAAAGTCTGGCTTTTGAAGACTGGATTCATGACCACATACATCTCGAAGGCAAGCCGATTCTTTTCCTTTGGAGAAATTCTCCTTCTGTCGTCATC-3′ This paper N/A
Recombinant DNA
pLOX-TERT-iresTK Addgene 12245
pLenti-EF1a-C-Myc-DDK-IRES-Puro OriGene PS10085
pLenti-EF1a-C-Myc-DDK-IRES-Puro-LIPT1-WT This paper N/A
pLenti-EF1a-C-Myc-DDK-IRES-Puro-LIPT1-N44S This paper N/A
pLenti-EF1a-C-Myc-DDK-IRES-Puro-LIPT1-S292X This paper N/A
Software and Algorithms
MultiQuant software v2.1 Applied Biosystems N/A
SIMCA-P v13.0.1 Umetrics N/A
Heatmap3 https://cran.r-project.org/web/packages/heatmap3/index.html N/A
MetaboAnalyst 3.0 http://www.metaboanalyst.ca/ N/A
HOPE-CMBI http://www.cmbi.ru.nl/hope/ N/A
PyMol https://pymol.org/2/ N/A