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The complex was concentrated to 0.5 ml using Ultra Centrifugal Filter (ThermoFisher MWCO, 100 kDa) and subjected to size-exclusion chromatography on a Superdex 200 Increase 10/300 column (GE Healthcare) that was pre-equilibrated with 20 mM HEPES pH 7.5, 100 mM NaCl, 2 mM MgCl 2 , 5 M ligands, 0.00075%(w/v) LMNG, 000025% glyco-diosgenin (GDN, Anatrace) and 0.00015% (w/v) CHS to separate complex from contaminants

This mislocalization increases cytosolic PUFA esterification, generating lipid peroxides that promote ferroptosis in neighboring epithelial cells, reinforcing the immune-epithelial crosstalk within the axis