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CBS proteins with a permanent C-terminal His-tag were purified using TALON (Clontech) resin, followed by anion exchange using a Hitrap Q column (Cytiva)

CBS deficiency led to markedly homocysteine accumulation and severely decreased levels of cystathionine and cysteine, which was due to impaired homocysteine removal through the transsulfuration pathway [32]

Results were analyzed using the 2 CT method, and the amount of target RNA was adjusted to the geometric mean of the five internal controls as previously described (Livak and Schmittgen, 2001)

Human studies have shown that as the severity of liver fibrosis progresses, the number of hepatic progenitor cells increase, mimicking the pathological processes associated with the wound healing and carcinogenic response of the liver to injury which is observed in chronic viral hepatitis and alcohol-related liver disease [91, 92]