Cells were grown onto 25 cm2 flasks and treated for 12 with vehicle (control) or 5 µM CoQ and then incubated for 24 h in fresh medium containing 5 µM Aβ25–35 peptide. Cells were detached, rinsed twice in PBS and homogenized by sonication in deuterated D2O (99.98% atom % D) (Sigma-Aldrich). Extracts were lyophilized and resuspended again in 40 µl of deuterated water (99.994 atom % D) (Sigma-Aldrich). NMR spectra were performed on a Varian VNMRS-400 NMR system using a nanoprobe with rotors of 40 µl. 1D and 2D NMR spectra were recorded for peak identification. The NMR spectra were processed with Mestrenova (Mestrelab Research) and peaks were also identified with Chenomx Profiler (Chenomx Inc.). Clustering and paired analysis for changes in the metabolites profile were assessed with the free software MEV 4.9 (http://www.tm4.org/mev.html ). Pathways enrichment was analyzed using the platform MetaboAnalist 2.0 (http://www.metaboanalyst.ca/MetaboAnalyst/faces/Home.jsp ).
Deuterated d2o
Manufactured by Merck Group
Deuterated D2O is a heavy water compound where the hydrogen atoms have been replaced with deuterium, a stable isotope of hydrogen. It is used as a specialized laboratory reagent for various analytical techniques, such as nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry.
Automatically generated - may contain errors
2 protocols using deuterated d2o
1
Metabolic profiling of Aβ25-35-induced cell damage
2
Graphene Oxide and Flake Treatment
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Cells were grown in 3 × 175 cm 2 flasks and treated for 7 days with 5 μg mL -1 GO or FLG. Cells were detached, rinsed twice in PBS and homogenized by sonication in deuterated D 2 O (99.98 atom% D) (Sigma-Aldrich) (n = 6).
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