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Subsequently, 200 µL FMOC-reagent (30 mM FMOC-Cl in acetonitrile) was added, and the reaction was gently mixed and incubated for 5 min

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Unlike the MTHFD1 encoded enzyme that is a tri-functional enzyme harboring N 5 , N 10 -methenyl-THF (5,10-methenyl-THF) cyclohydrolase, 5,10-methylene-THF dehydrogenase, and 10-formyl-THF synthetase activities, the MTHFD1L encoded enzyme only possesses 10-formyl-THF synthetase activity