Gene expression for eNOS S1176 mutant mice

Extensive reports in the past decade have shown the importance of eNOS expression in atherogenesis. However, both the eNOS knockout and overexpression mouse models exhibit paradoxical results of increased atherosclerotic lesions. Our genetic approach herein utilizes endogenous point-mutations at the eNOS S1176 site to effectively manipulate eNOS activity rather than total eNOS protein expression levels. Our efforts definitively show that eNOS S1176 phosphorylation is sufficient to mitigate atherosclerotic lesion progression. eNOS S1176A ‘loss-of-function’ and eNOS S1176D ‘gain-of-function’ mice were fed either a standard laboratory diet or a high-fat, high-cholesterol Western Diet for 4weeks. Whole aortas were isolated (2-3 mice per group) for total RNA extraction and quality checks. Satisfactory samples were prepared for hybridization reactions with the NanoString platform (PanCancer Immune Profile Mouse). Each sample reflects a single aorta. mRNA profiling reflects 770 target genes including 20 housekeeping genes, 8 negative and 6 positive spike-in controls.