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SDSPAGE and western blotting Samples were fractioned by SDSPAGE on 8% gels under reducing conditions, immunoblotted onto nitrocellulose membranes (GE Healthcare) and bands detected by enhanced chemiluminescence (GE Healthcare) onto films developed on a Xograph Compact X5 processor

4.5.2 ER stress-induced CHAC1 expression and ferroptosis ER stress-induced CHAC1 expression contributes to ferroptosis by disrupting GSH homeostasis, which is crucial for cellular antioxidant defense

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Mechanistically, a microglial inflammation-oxidative stress feedforward loop perpetuates injury: Activated microglia secrete neurotoxins, impairing neuronal mitochondrial function and antioxidant capacity (such as GPX4), thus increasing ROS leakage