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The following factors were used: 50 ng/mL recombinant EGF, 500 nM A83-01 (Biogems), 100 ng/ mL recombinant Noggin (Peprotech), 10 M Y-27632 (Abmole), 500ng/mL R-spondin-1(Peprotech), 10 mM nicotinamide (Sigma), and 10 M SB202190 (Sigma)

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Two gut microbiota inocula from the feces of high-TMAO producers were prepared by mixing 1 g of feces with 15 mL of PBS with 0.05% cysteine and filtering the mixture through a 100-m Falcon cell strainer, following which 100 L of the stool filtrate was transferred to 1.9 mL of medium and subsequently incubated at 37 C under anaerobic conditions (80% N 2 , 10% CO 2 , and 10% H 2 ) for 0, 6, 12, and 24 h