Fig. 1

Experimental design and bioinformatics workflow. A. Human pulmonary microvascular endothelial cells (HPMEC) and human lung epithelial cells (BEAS-2B) were incubated with DMEM + 10% FBS (D10) at 37 °C until sub-confluent, and then preserved with lung preservation Perfadex^® solution at 4 °C with 50% O₂ for 18 h cold ischemic time (CIT) to simulate static cold storage of donor lung. Cells were then switched to either D10 to simulate reperfusion during lung transplantation, or with Steen solution to simulate ex vivo lung perfusion (EVLP), respectively, for 4 h at 37 °C. B. Total RNA was extracted after CIT 18 h, reperfusion 4 h, or EVLP 4 h. After purity check, RNA samples were sequenced, and raw data (FASTQ files) were processed as indicated in the flow chart for differentially expressed gene (DEG) analysis and gene set enrichment analysis (GSEA)