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PCs ( P11) were seeded into collagen type IV and fibronectin coated dishes in Dulbeccos modified Eagles medium (DMEM, Life Technologies, Thermo Fisher Scientific, USA) supplemented with 20% FBS, 1% Glutamax (Life Technologies) and gentamicin (50 g/ml)

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In addition to acetyl-CoA, there are three other pathways known to produce butyrate, glutarate, 4-aminobutyrate, and lysine and all pathways coalesce at a central energy-generating step where crotonyl-CoA is transformed into butyryl-CoA, catalyzed by the electron-transferring flavoprotein butyryl-CoA dehydrogenase (52)