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The pellet was resuspended in 800 l 0.2 M sulphuric acid and incubated on a roller for 2 h at 4C, followed by centrifugation for 10 min at 11,000 g to remove cellular debris

Using membrane-binding assays, we found that replacing residues 157 to 176aa of the AH with a flexible linker (GGGSGGGSGGGS, AH-mutant) significantly reduced AtSNX1s ability to bind liposomes, thereby impairing its membrane tubulation capacity (Fig

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