The tryptic peptides were fractionated into fractions by high-pH reversed-phase HPLC using a Thermo Betasil C18 column (5-μm particles, 10 mm ID, 250 mm length). Briefly, peptides were first separated with a gradient of 8 to 32% acetonitrile (pH 9.0) over 60 min into 60 fractions. Then, the peptides were combined into 10 fractions and dried by vacuum centrifugation.

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