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<p>Subcuteaneous WAT was later shattered in liquid nitrogen, and around 100 mg was taken for preparation. All ~5 animals per cohort had their RNA prepared, and then were pooled evenly (by µg of RNA) into a single RNA sample for each cohort. The pooled RNA samples were then purified using RNEasy, then sent out for array analysis. All RIN values were > 8.0. RNA was prepared in the summer of 2013, while the RNEasy cleanup occurred in the winter of 2015; unlike the other tissues run as a part of this study, these RNA samples spent a significant time in the -80° freezers. However, the samples only underwent one freeze—thaw cycle.</p>
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