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Reverse Transcription-Quantitative Polymerase Chain Reaction The total RNA content was isolated from cells or the tumor using TRIzol reagent (Invitrogen), treated with DNase I (Thermo Fisher Scientific, Cat# EN0521), and then reverse transcribed into cDNA using a Maximal First Strand cDNA Synthesis Kit (Thermo Fisher Scientific, Cat# K1622)

Next, glutamine is converted into glutamate by GLSs, amidohydrolase enzymes that catalyze the conversion of glutamine into glutamate, releasing ammonium ions

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Some evidence suggests that AOD-9604 may have effects on the heart even beyond its capacity to reduce fat burden