Before liquid chromatography–tandem MS (LC-MS/MS) analysis, each OFFGEL peptide fraction was reconstituted in ddH20/ACN/formic acid (50:49.9:0.1) and then shaken at 37°C and vortexed thoroughly. Chromatographic separation and mass spectra acquisition (LTQ Orbitrap Velos Pro, Thermo Fisher Scientific) was performed on each fraction and was previously described (17), with a modified MS1 resolution of 60,000 and MS2 resolution of 30,000.

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