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Samples were loaded onto the gel filtration column and eluted isocratically in 100 mM sodium phosphate + 100 mM NaCl buffer pH 7.2 at a flow rate of 0.5 ml min−1 for 50 min. The elution was monitored with a Ultimate 3000 Photodiode Array detector (Dionex) coupled with the Ultimate 3000 HPLC system (Dionex).

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The LC/MS instrument consisted of a Dionex Ultimate 3000 Rapid Separation UPLC system equipped with a Dionex Ultimate 3000 autosampler and a Dionex Ultimate 3000 photodiode array detector coupled with a Bruker MicrOTOF-Q II quadrupole time-of-flight hybrid mass spectrometer using Hystar 3.2 software.

Twenty per cent of the peptide mixture was analysed by nano-liquid chromatography tandem mass spectrometry using an LTQ Velos-Orbitrap MS (Thermo Scoupledic) coupled with an Ultimate RSLCnano-LC system (Dionex).

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Peptides were analysed with on-line nanoLC-MS/MS on a LTQ FT mass spectrometer (ThermoElectron) coupled with an Ultimate 3000 Nano/Capillary HPLC System (Dionex).

Generated peptides were analysed online by nano-flow HPLC-nanoelectrospray ionization using a LTQ Orbitrap XL mass spectrometer (Thermo Fisher Scientific) coupled with an Ultimate 3000 HPLC (Dionex).

The differentially labelled samples were mixed correspondingly before the nanoLC-MS/MS analysis on a LTQ Orbitrap Velos (Thermo Fisher) hybrid mass spectrometer equipped with a nanospray source, coupled with an Ultimate 3000 RSLCnano System (Dionex).

Peptides were redissolved in 0.5% formic acid and analyzed with online nanoLC-MS/MS on a LTQ FT mass spectrometer (Thermo Fisher Scientific) coupled with an Ultimate 3000 Nano/Capillary LC System (Dionex).

Briefly, TMT labelled and fractionated samples (~1 µg) were loaded onto Liquid chromatography-MS/MS analysis, using Orbitrap Fusion Lumos Mass Spectrometer coupled with a Dionex Ultimate 3000 nano-LC pump.

Analysis of peptides was performed on a Velos orbitrap (Thermo Scientific, Bremen, Germany) mass spectrometer coupled with a Dionex Ultimate 3000 RS (Thermo Scientific).