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15,16 REM sleep concludes with a return to N1, sometimes with a brief period of wakefulness prior to the next sleep cycle

Sample preparation for serum metabolite and untargeted/targeted metabolomic mass spectrometry methods performed in this study were based on previously established protocol (Figure 1

10.1016/j.metabol.2009.05.014 67

Plates were centrifuged at 900 g for 90 min at 33 C (spinfection)

3.2.2 Dynamic monitoring of glutamine metabolism by extracellular metabolite quantification assays To monitor the metabolic activity of a pathway, the real-time viability assay is one of the most widely used tools, which measures the rate of uptake/excretion of metabolites such as glucose, glutamine, lactate, and ammonia, providing an overview of overall cellular metabolism (Antoniewicz, 2013a