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Cell viability measurements For this experiment, approximately 12,000 senescent and 6,000 non-senescent cells were seeded in triplicate wells of 96-well plates

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Hyperactivation of oncogenes leads to excessive replication stress and activation of the DDR pathway.15 Oxidative Stress and Mitochondrial Dysfunction: Reactive oxygen species (ROS) generated from mitochondrial dysfunction or environmental stress contribute to cellular senescence.16 Oxidative stress leads to DNA damage, lipid peroxidation, and protein dysfunction.17 ROS can activate the p38 MAPK pathway, which enhances p53 and p16 signaling, reinforcing senescence.18 Importantly, oxidative stress is not an isolated mechanism but a central upstream driver of the SASP

EVIDENCE GRADES Original Schoenenberger 1977 isolation paper described EEG-pattern shift toward slow-wave sleep in rabbits