title: GSM5825617: CD206neg.IM.WT_R1; Mus musculus; RNA-Seq
library_construction_protocol: For bulk mRNA-sequencing (RNA-seq), lungs from IM-depleted and control mice were isolated, chopped and incubated with collagenase I and DNaseI at 37 degrees for 30 minutes. Mononuclear cells were released and purified by gredient centrifugation with Percoll. Alveoloar macrophages (AM), CD206+ interstitial macrophages (CD206+ IM) and CD206- IM were FACS-sorted into Trizol. Total RNA was extracted with standard Trizol RNA extraction protocole. RNA quality and quantity were evaluated using a 2100 bioanalyzer (Agilent) and the Quant-iT™ RiboGreen™ RNA Assay Kit (ThermoFisher). One hundred nano-grams of RNA was used to generate the libraries using the Truseq stranded mRNA kit (Illumina). These libraries were sequenced on an Illumina Novaseq sequencer on a SP flow cell.