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The organic layer was reconstituted in chloroform:methanol (2:1 v/v, 200 l and diluted 1:10 with 2:1:1 isopropanol:acetonitrile:water, with the addition of an internal standard mix composed of isotopically labelled compounds from major lipid classes
MOTS-c's AMPK activation aligns with the body's metabolic circadian rhythm, which peaks in the morning and early afternoon
The cells were then lysed in 200 L 1 RIPA buffer + 50 mM IAA and incubated on ice for 30 min followed by a snap freeze
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