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Methyltransferase activity of NNMT
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To analyze the methyltransferase activity of NNMT, we used an assay based on the conversion of nicotinamide to radioactively labeled N-methylnicotinamide with 14C-Ado-Met as the methyl donor as previously described65. Cell lysates were prepared from UC7 and UC11 cell lines with shRNA silenced AURKA and NNMT were incubated with Nicotinamide and 14C-Ado-Met. Lysates incubated with DMSO served as controls. The radioactivity was measured using a Beckman liquid scintillation counter.
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