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The fluorescence intensity of the ROS in the cells was detected with the use of a fluorescent enzyme marker and a fluorescence light microscope

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Importantly, these changes in mitochondrial mass are not related to engagement of apoptosis: inhibition of caspase activity using zVAD-fmk did not prevent increases in mitochondrial abundance per cell (Extended Data Fig

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These results directly confirm that excessive NNMT activation suppresses NAD+ synthesis and impairs energy metabolism, while NNMT inhibition effectively reverses this process, establishing NNMT as a negative regulator of energy metabolism