MetaboLights

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ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14692/m_MTBLS14692_LC-MS_alternating_reverse-phase_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14692/s_MTBLS14692.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14692/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14692/a_MTBLS14692_LC-MS_alternating_reverse-phase.txtprimary200OKftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14692Lipid annotation was performed in MS-DIAL based on accurate mass and MS/MS spectral matching. Only features with MS/MS available were retained for downstream analysis. The adducts considered for annotation were restricted to [M−H]− and [M+CH3COO]− in negative ionization mode, and [M+H]+, [M+Na]+, and [M+NH4]+ in positive ionization mode.MetaboLightsPublicLiquid Chromatography MS - alternating - reverse-phaseLipid extracts were analyzed by UHPLC–MS/MS in both positive and negative ionization modes. Chromatographic separation was performed on an Accucore™ C30 column (150 × 2.1 mm, 2.6 μm; Thermo Fisher Scientific, Waltham, MA, USA) using a Vanquish™ UHPLC system (Thermo Fisher Scientific). Separation was conducted using a 20-min gradient at a flow rate of 0.35 mL/min and a column temperature of 40 °C. Mobile phase A consisted of acetonitrile/water (6:4, v/v) with 10 mM ammonium acetate and 0.1% formic acid, and mobile phase B consisted of acetonitrile/isopropanol (1:9, v/v) with 10 mM ammonium acetate and 0.1% formic acid.Differential control of brown adipose tissue function by dietary and Scd-derived MUFAs reveals a non-redundant role of Scd2.TINGXUAN GAONanjing agricultural universityCell pelletmass spectrometry assay Briefly, 0.75 mL methanol was added to each 100 mg tissue or 106 cells in a centrifugation tube with a Teflon-lined cap and vortexed thoroughly. Then, 2.5 mL MTBE was added and the mixture was incubated for 1 h at room temperature with shaking. Phase separation was induced by adding 0.625 mL MS-grade water. After 10 min incubation at room temperature, samples were centrifuged at 1,000 × g for 10 min. The upper (organic) phase was collected, and the lower phase was re-extracted with 1 mL MTBE/methanol/water (10:3:2.5, v/v/v). The combined organic phases were dried and reconstituted in 100 μL isopropanol prior to LC–MS/MS analysis. A pooled QC sample was prepared by combining equal aliquots from each biological sample and was processed in parallel with biological samples Mus musculushttps://www.ebi.ac.uk/metabolights/MTBLS14692TINGXUAN GAO. Nanjing agricultural university. 2022208014@stu.njau.edu.cn.Raw LC–MS/MS files were processed using MS-DIAL for peak detection, MS/MS deconvolution, alignment, and lipid annotation. Centroid mass tolerances were set to 0.01 Da for MS1 and 0.025 Da for MS2. Peak detection was performed with a minimum peak height of 1000 amplitude and a mass slice width of 0.1 Da. Smoothing was applied using a linear weighted moving average with a smoothing level of 3 scans, and the minimum peak width was set to 5 scans. MS/MS deconvolution was conducted using a sigma window value of 0.5, an MS/MS abundance cutoff of 10 amplitude, and an MS/MS relative abundance cutoff of 1%. Only features with MS/MS available were retained. The exported feature table was further curated by applying a fill rate cutoff >0.6 across all samples and reducing redundant entries derived from multiple adducts or closely overlapping aligned peaks. One representative feature was retained from each redundancy group based on annotation confidence and signal robustness.Oleic acid supplementBAT differentiationSCD1/2 knock out2022208014@stu.njau.edu.cnBrown adipose tissue was rapidly collected from mice after sacrifice. The interscapular brown adipose tissue depot was carefully dissected, cleaned of surrounding connective tissue and visible white adipose tissue, and immediately snap-frozen in liquid nitrogen. Frozen samples were stored at −80°C until lipid extraction and LC-MS/MS-based lipidomic analysis.MetabolomicsThermo Scientific Vanquish UHPLC SystemC2C12Mus musculusThermo Scientific Q Exactiveuntargeted analysisstearoyl-CoA desturasebrown adipose tissue differentiationCell pelletLipidomicsexperimental blankThermo Scientific Vanquish UHPLC SystemC2C12Mus musculusThermo Scientific Q Exactiveuntargeted analysisstearoyl-CoA desturasebrown adipose tissue differentiationCell pelletLipidomicsexperimental blankThe UHPLC system was coupled with a Q Exactive™ HF/Q Exactive™ HFX mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA). Mass spectrometry was performed in both positive and negative ionization modes. The MS parameters were as follows: sheath gas 40 psi, auxiliary gas 10 L/min in positive mode and 7 L/min in negative mode, spray voltage ±3.5 kV, capillary temperature 320 °C, heater temperature 350 °C, S-lens RF 50, scan range m/z 144–1700, AGC target 3e6 for MS1 and 2e5 for MS2, maximum injection time 100 ms, isolation window 1 m/z, normalized collision energy 22/24/28 eV, and dynamic exclusion 6 s.falseLipidomic profiling of Scd1- and Scd2-deficient C2C12 cells during brown adipogenic differentiation with oleic acid supplementationThis study investigated the lipidomic remodeling of Scd1- and Scd2-deficient C2C12 cells during brown adipogenic differentiation. Scd1 or Scd2 knockout cells and corresponding control cells were induced to undergo brown adipocyte-like differentiation for 96 h. To evaluate whether exogenous monounsaturated fatty acid supplementation could rescue lipid metabolic alterations caused by Scd1 or Scd2 deficiency, cells were supplemented with 10 μM oleic acid during differentiation. Lipid extracts were analyzed by LC-MS-based lipidomics to characterize changes in lipid classes and molecular species associated with Scd1 or Scd2 loss and oleic acid-mediated rescue during the differentiation process.2026-06-232026-06-06MTBLS14692