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ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/m_MTBLS12815_NMR___metabolite_profiling_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/a_MTBLS12815_NMR___metabolite_profiling.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/s_MTBLS12815.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/3-M8.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/1-C4.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/1-M4.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/3-C8.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/6-M9.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/6-C10.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/1-C3.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/6-M12.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/6-M11.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/3-M5.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/3-C7.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/6-C11.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/1-C2.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/3-C6.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/1-M2.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/1-M1.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/6-M10.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/3-M6.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/1-C1.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/Acquisition.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/6-C12.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/3-C5.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/1-M3.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/6-C9.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815/FILES/RAW_FILES/3-M7.zipprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12815Metabolitesand all identified molecules were quantified using Chenomx NMR suite (Chenomx Inc.)), with the TSP peak at 0.00 ppm serving as the chemical shift and concentration reference. The absolute concentration of TSP was calibrated using the ERETIC2 method (TopSpin 3.5, Bruker), based on an in-house reference sample of dimethyl sulfone (Sigma-Aldrich) prepared in the same buffer as the test samples. Metabolite concentrations (µM) were normalized to the protein mass (mg) of each extract. Protein content was determined using the Pierce BCA Protein Assay Kit (Thermo Fisher Scientific), following the manufacturer’s instructions.MetaboLightsPublicNuclear Magnetic Resonance (NMR) -MTCH2 modulates CPT1 activity to regulate lipid metabolism of adipocytes.1H-NMR spectradata were acquiredobtained using a Bruker 600 MHz AVANCE II 600 MHz spectrometer equipped with a 5 mm TCI cryo probe, employing the noesygppr1d pulse sequence (TopSpin 3.5, Bruker).Chunyan WuETH zurichAdipocytesNMR spectroscopyIntracellular metabolites were then extracted using a pre-chilled mixture (-80 degree) of methanol/chloroform/water, 6.75/0.75/2.5. The cellular extracts were collected and incubated on dry ice for 30 min. Subsequently, the extracts were centrifuged at 0 degree and 18000 xg for 15 min. The supernatants were collected and dried using a stream of nitrogen gas. All samples were maintained on dry ice during processing. Media samples were collected at the specific time points and mixed with 2 volumes of LC-grade methanol (Sigma Aldrich). Homo sapienshttps://www.ebi.ac.uk/metabolights/MTBLS12815Chunyan Wu. ETH Zurich. chunyan.wu@hest.ethz.ch.Christian Wolfrum. ETH Zurich. christian-wolfrum@ethz.ch.Metabolitesand all identified molecules were quantified using Chenomx NMR suite (Chenomx Inc.)), with the TSP peak at 0.00 ppm serving as the chemical shift and concentration reference. The absolute concentration of TSP was calibrated using the ERETIC2 method (TopSpin 3.5, Bruker), based on an in-house reference sample of dimethyl sulfone (Sigma-Aldrich) prepared in the same buffer as the test samples.MTCH2 knockdownchunyan.wu@hest.ethz.chDifferentiated hMADS adipocytes on day 15 were transfected with control or MTCH2 siRNA pools (Microsynth) using Lipofectamine RNAiMAX (Invitrogen), following manufacturer’s instructions. A control siRNA pool was used as a control. Following a 24 hour transfection period, the medium was replaced with fresh adipogenic medium containing 100nM rosiglitazone. Adipocytes were then treated with cAMP for 1hr, 3hr and 6 hr on day 18, at which point adipocytes were harvested for cellular metabolites measurements between control and MTCH2 KD groups (n=4 biological replicates per group per time point, 24 samples in total). Differentiated adipocytes cells were quenched with liquid nitrogen after washing with ice-cold PBS. 1H-NMR spectradata were acquiredobtained using a Bruker 600 MHz AVANCE II 600 MHz spectrometer equipped with a 5 mm TCI cryo probe, employing the noesygppr1d pulse sequence (TopSpin 3.5, Bruker).Metabolomicsglucosenuclear magnetic resonance spectroscopytargeted metabolitesglucosenuclear magnetic resonance spectroscopytargeted metabolitesThe dried extracts were reconstituted with in 150 mM potassium phosphate buffer (0.15 M K2HPO4, pH 7.04) in prepared in deuterium oxide (D2O) deuterated water for NMR analysis. A fixed amount of trimethylsilyl propionate-d4 sodium salt (TSP-d4, Cambridge Isotope Laboratories) was added to each sample as an internal standard (0.05 mM for cell extracts) to enable quantification.L-glutamic acid2-oxoglutaric acidsuccinic acidfalseMTCH2 modulates CPT1 activity to regulate lipid metabolism of adipocytesTo understand the molecular changes that underwrite enhanced mitochondrial respiration upon MTCH2 knockdown, we performed lipidomics profiling in hMADs adipocytes.2025-08-042025-08-04MTBLS12815MTBLC30915MTBLC16015MTBLC15741CHEBI:30915CHEBI:16015CHEBI:15741