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For the NAC rescue experiment, mice received intraperitoneal administration of 200 mg/kg of body weight NAC (A105422, Aladdin) in phosphate-buffered saline (pH 7.4) every second day for two weeks

Cells were harvested in 200 L PBS + 0.1% BSA and subjected to flow cytometry analysis to examine oxidation of BODIPY 581/591 C11, a reporter for lipid ROS levels in cells

When to use SUMO: When you need a native N-terminus (structural biology, functional studies) When tag size matters but you still want solubility enhancement When clean, efficient cleavage is critical Excellent for proteins destined for crystallization or NMR When NOT to use SUMO: In eukaryotic expression systems (host SUMO proteases will cleave prematurely) When the N-terminus is buried or critical for folding (Ulp1 won't access it) GST (Glutathione S-Transferase) Size: ~26 kDa (211 residues) Mechanism: GST provides moderate solubility enhancement and convenient purification via glutathione-Sepharose

Nrf2 regulates important genes such as heme-oxygenase 1(HO-1), glutathione S-transferase (GST), glutathione reductase (GR), glutamate-cysteine ligase subunits (GCLc and GCLm), NAD(P)H quinone oxidoreductase-1 (NQO1), and thioredoxins (TrxR1), among others