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.