Illumina NovaSeq 6000 paired end sequencing; RNA-seq of ex vivo cultured lung CD140a+ mesenchymal cells (fibroblasts) upon stimulation by IL-1β
RNA-seq of ex vivo cultured lung CD140a+ mesenchymal cells (fibroblasts) upon stimulation by IL-1β
Sample 6_p
Mice were euthanized and lungs were harvested and digested. Lung single cell suspensions were prepared and CD45-CD31-CD326-CD140a+ cells were sorted, and ex vivo cultured for further experiments. Ex vivo cultured lung CD140a+ mesenchymal cells (fibroblasts) were stimulated with vehicle or IL-1β for 24 hours.  Total RNA was isolated from lung CD140a+ mesenchymal cells (fibroblasts) using the miRNeasy Mini kit. Stranded libraries were prepared using the KAPA mRNA HyperPrep Kit: isolation of polyA containing mRNA using oligo-dT magnetic beads, RNA fragmentation, first and second strand cDNA synthesis, ligation of Illumina-specific adapters containing a unique barcode sequence for each library, and PCR amplification. Libraries were checked for quality and concentration using the DNA 1000 assay and quantitative PCR, following the manufacturers' instructions.