2.12 Proteomics analysis
Lung tissues from mice and cells from the chips were harvested for
proteomic studies. Tissues and cells were lysed with SDT buffer and
sonicated intermittently (10 seconds on, 10 seconds off) for a total of
2 minutes to enhance disruption. Centrifugation at 12,000 rpm for 10
minutes ensued to discard insoluble fragments. Proteins were extracted
and subsequently digested using the filter-aided sample preparation
(FASP) method. Protein alkylation was accomplished using IAA, followed
by trypsin digestion to generate peptide fragments. Following peptide
desalting, data acquisition was conducted using an LC-MS/MS system
consisting of Thermo Fisher’s Easy-nano LC and QE Plus instruments.