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Following isolation and quantification, mitochondria were divided into at least 100 g replicates in microtubes and centrifuged at 11,800 g for 10 min to pellet and were used for respiration and NAD + uptake assays or flash-frozen for metabolite analyses, including NAD + , NADH and metabolomics

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The acquired genetic alterations confer upon the initially transformed atypical melanocytes and later upon the melanoma cells that evolved, a selective advantage over their neighbouring normal cells in terms of fitness and proliferation