Genetic rescue of mitochondrial complex I reverses metabolic defects in circadian mutant mice

We used single nuclei RNA-seq to compare gene expression from adipocyte circadian mutant mice +/- expression of the yeast NADH dehydrogenase (NDI1). We used RNA-sequencing of fractionated adipocytes from the visceral white adipose tissue of control mice, mice lacking the circadian clock activator Bmal1, and mice lacking the mitochondrial complex I subunit Ndufs2. Overall design: Visceral white adipose tissue (WAT) was isolated from 3-month old male mice and was homogenized, followed by filtering and centrifugation to isolate nuclei. Nuclei were then analyzed for single nuclei RNA-sequencing with 10X genomics. Bulk RNA-seq: Visceral white adipose tissue (WAT) was isolated from 3-month old male mice and was digested with collagenase followed by differential centrifugation to float the adipocytes. The adipocyte fraction was then homogenized followed by RNA isolation, library preparation, and sequencing.