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In addition, in Complex I, metformin directly down-regulates mitochondrial glycerol 3 phosphate dehydrogenase (mGPDH), leading to an increased cytosolic redox statewith a reduction in nicotinamide adenine dinucleotide reduced form (NADH) and increased nicotinamide adenine dinucleotide (NAD)to reduced gluconeogenesis from lactate, and to a decrease in the activity of the glycerolphosphate shuttle (which transfers NADH from the cytosol to mitochondria)

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Knudsen chose to use palmitic acid which is one of the most common fatty acids found in the body (Holst 2024