MTCH2 modulates CPT1 activity to regulate lipid metabolism of adipocytes
Description
To understand the molecular changes that underwrite enhanced mitochondrial respiration upon MTCH2 knockdown, we performed unbiased metabolomics and lipidomics profiling in hMADs adipocytes.
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Steps to reproduce
NMR spectra from cell extracts were imported in Chenomx for quantification. The IS in Chenomx is TSP and the concentrations of TSP per sample can be found in the uploaded excel file. ChenomX concentrations are then corrected for protein weight per sample. For cell media the process is describe below: Calculation of Metabolite Uptake and Release Uptake or release of extracellular metabolites was calculated using the following formula: Uptake/Release (nmol/mg protein) = (C_spent – C_fresh) / protein mass Where: • C_spent is the amount of metabolite in the spent (cell-containing) medium (in nmol), • C_fresh is the amount in fresh (cell-free) medium (in nmol), calculated as the average of 3 blank media samples incubated under the same conditions, • Protein mass (in mg) was determined for each sample. NMR concentrations (in µM) were converted to nmoles using the following equation: C (nmol) = [NMR concentration (µM) × 0.22 mL × 7 mL] / 0.3 mL Note: Glutamine values in the extracellular data may be affected by the presence of Glutamax (Gln-Ala) in the medium, which was not quantified.