Before each LC-MS/MS measurement, all peptide samples were desalted and purified on C18 StageTips (53). Reversed-phase C18 discs (Empore) were activated with methanol and equilibrated with solvent A*. Up to 10 μg of peptides was loaded onto the membrane and washed with solvent A. Peptides were eluted with 50 μl of solvent B [80% (v/v) acetonitrile and 0.1% (v/v) formic acid] and concentrated by vacuum centrifugation. The sample volume was adjusted with solvent A and final 10% (v/v) of solvent A*.

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