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Briefly, flash-frozen tissue samples were homogenized in ice-cold PBS containing 2 mM EDTA (1 ml PBS/EDTA per 10 mg tissue) using a Dounce homogenizer

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Decreased activity of the mitochondrial respiratory chain was reported in overweight/obese patients with NASH compared to controls [97, 98], and alterations in mitochondrial ultrastructure have been observed prior to NAFLD development in the Otsuka LongEvans Tokushima Fatty rat [99] as well as in patients with steatosis preceding progression to NASH [100]