Proteomic analysis of injured and uninjured nerves

Murine injured sciatic nerves were harvested from C57BL/6J mice at 24h post-transection. Uninjured nerves were set as control group. Nerve samples were frozen and then ground into fine and uniform powder in liquid nitrogen for protein extraction. Then the samples were digested and fractionated. The data independent acquisition (DIA) mass spectrometry analysis was acquired by TimsTOF Pro2 mass spectrometer (Bruker) and nanoElute (Bruker). For data independent acquisition (DIA), 56 DIA windows were acquired and the collision energy was ramped linearly as a function of the mobility from 59 eV at 1/K0 = 1.6 Vs/cm2 to 20 eV at 1/K0 = 0.6 Vs/cm2. The MS/MS spectra were recorded from 100 to 1700 m/z. The default factory settings were used for the Spectronaut Pulsar™ 15.3.210906.50606 (Biognosys, Swiss) search and library generation.