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Western blotting Tissue lysates were prepared with radioimmunoprecipitation assay buffer and separated on NuPAGE Bis-Tris Mini Gels (Life Technologies), followed by the transfer to polyvinylidene difluoride membrane (Millipore) following a standard protocol

Recombinant pure human GCLM (0.1 g), provided by from Ono Pharmaceuticals, was pre-treated with DMSO or the covalently-acting EN25-alkyne probe (10-100M final concentration) for 30 minutes at 37 C in a volume of 50L of PBS

the present invention provides a composition for treating hepatic steatosis, nonalcoholic steatohepatitis (NASH), fibrosis, type-2 diabetes mellitus, obesity, hyperlipidemia, hypercholesterolemia, atherosclerosis, cognitive decline, dementia, cardiorenal diseases such as chronic kidney diseases or heart failure comprising an acceptable carrier and a compound of Formula I, or a pharmaceutically acceptable salt thereof

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Nat Commun 10(1):112 Bahri S, Curis E, El Wafi FZ, Aussel C, Chaumeil JC, Cynober L, Zerrouk N (2008) Mechanisms and kinetics of citrulline uptake in a model of human intestinal epithelial cells