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(4) Centrifuge for 10 min (12000 rpm, 4C), aspirate 150 L of the supernatant with a syringe, filter it using a 0.22 m organic phase pinhole filter, transfer it to an LC injection vial and store it at -80C until LC-MS analysis

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Isolation of WAT stromal vascular cells and adipocyte differentiation To isolate the stromal vascular fraction, minced iWAT (combined depots from 45 mice per genotype) from 46-week-old mice was digested in 1 mg ml 1 collagenase D (Roche) with 1.5% BSA in sterile Hanks Balanced Salt Solution and incubated in a 37 C shaking water bath for 1 h