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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)

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DMT-1 is expressed at the brain endothelial and functions as a high-affinity transporter for non-transferrin-bound divalent cations, exhibiting a non-selective affinity for various divalent metal ions, including Mn [15, 133]

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)

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