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Plasma, liver, and epididymal fat pad samples were harvested for analysis

Methods Reagents and antibodies Anti-Acsl4 antibody (ab155282, abcam), Anti-Fsp1 antibody (68049-1-Ig, Proteintech), Anti-Keap1 antibody (ab227828, Abcam), Anti-Gpx4 antibody (67763-1-Ig, Proteintech), Anti-F4/80 antibody (ab6640, Abcam), Anti- Ly6G antibody (ab25377, Abcam), Anti-Megalin antibody (ab184676, Aabcam), Anti-4-HNE antibody (ab48506, Abcam), Anti-GAPDH antibody (60004-1, Proteintech), BODIPY 581/591 C11 lipid peroxidation probe (D3861, Invitrogen), Mito-Tracker Red CMXRos fluorescent probe (C1035, Beyotime Biotechnology)

These improved biomarkers correspond with endpoints such as improved gait and body composition

A consistent bedtime routine can further enhance its benefits