2.1 Chemicals and reagents
Acetonitrile (ACN, MS grade), trifluoroacetic acid (TFA, MS grade),
formic acid (FA, MS grade), PierceTM BCA Protein Assay Kit and HeLa
Protein Digest Standard were purchased from Thermo Scientific (St.
Louis, MO, USA). Culture media such as Dulbecco’s Modified Eagle’s
Medium (DMEM), penicillin/streptomycin (P/S), fetal bovine serum (FBS)
and trypsin-EDTA were from Gibco Life Technologies (New York, USA). The
iST kit for proteomic sample preparation was purchased from PreOmics
GmbH (Planegg, Germany). Deionized water used in the experiments was
prepared by the Milli-Q50SP system (Millipore, Milford, MA).
2.2.
Patients and sample collection
The present study was conducted in accordance with the guidelines of the
Declaration of Helsinki with the approval of the Research Ethics
Committee from
Ruijin
Hospital, Shanghai Jiaotong University School of Medicine
(1.0/2019-10-1).
Written
informed consent was obtained from all patients or their legal
representatives prior to their participation in the study.
A total of 29 samples from gliomas and BrMs patients were obtained from
the Ruijin Hospital, Shanghai Jiao Tong University School of Medicine
between October 2020 and September 2021. Details of clinical information
on the subjects were shown in Figure 2A and Table S1. Tumor tissues were
collected after surgery, and were washed three times with PBS. After
that, tissues were immediately transferred to liquid nitrogen and
collected into 2 mL cryogenic storage vials (Corning, New York, USA),
and stored at −80 °C for later use.