Single cell RNA sequencing on rats with or without bleomycin treatemnt
Purpose:To characterize the heterogeneity of endothelial cells in bleomycin-induced lung fibrosis in rats Overall design: After adequate anesthesia, BLMwas administered by oral tracheal intubation via a 16-gauge catheter. One dose of BLM (5 mg/kg, the BLM-treated group) or an equal volume of saline was administered via the catheter. The rats were rotated immediately after instillation to ensure thorough drug distribution in the lungs. All rats were killed by cervical dislocation and lungs were harvested 28 days after intratracheal BLM or saline treatment. Rat lungs were harvested and 3 mm3 lung tissues were taken for digestion to generate single-cell suspensions.Beads with unique molecular identifier (UMI) and cell barcodes were loaded close to saturation, so that each cell was paired with a bead in a microwell. Whole transcriptome libraries were prepared using the BD Rhapsody system (BD Bio_x0002_sciences).