Effect of the combined neonatal pneumonia virus of mice infection and house dust mite exposure on the trancriptional profile of dendritic cells in the lung
Effect of the combined neonatal pneumonia virus of mice infection and house dust mite exposure on the trancriptional profile of dendritic cells in the lung
Single cell suspensions of the lung were obtained and cells were stained for 30 minutes at 4 degrees celcius before sorting. Following sorting strategy was used: live cells, CD45 IV negative cells. For each sample, 7500 immune cells were sorted (CD45+) and 7500 structural cells (CD45-, CD31-, Ter119-, EPCAM+) Single live cells were sorted and loaded onto a Chromium GemCode Single Cell Instrument (10x Genomics) to generate single-cell gel beads in emulsion (GEMs) The scRNA-Seq libraries were prepared using the GemCode Single Cell 3' Gel Bead and Library kit, version NextGEM 3.1 (10x Genomics) according to the manufacturer's instructions