GSM8774328: ILC2, mes ln, infected 2; Mus musculus; RNA-Seq
Intestine draining mesenteric lymph nodes, lung, and bone marrow were collected 5 days post infection with Nippostrongylus brasiliensis. ILC2 and ILC progenitors were enriched using EasySep™ Mouse Hematopoietic Progenitor Cell Isolation Kit (Stemcell, 19856) prior to cell sorting. Bone marrow cells were sorted for progenitor cell markers CD45+Lin- CD127+ a4b7+ Flt3-. These markers account for both the common helper-like ILC progenitors (CHILP) and ILC2 progenitors (ILC2p). Mesenteric lymph nodes and lung cells were sorted for conventional ILC2 markers CD45+ Lin- CD127+ and KLRG1low/+. Lung, mesenteric lymph node, and bone marrow cell suspensions from three mice were treated as 9 separate samples for scRNA-seq. Sorted cells were resuspended in 0.04% BSA in PBS and counted prior to GEM encapsulation using the 10x Genomics Chromium controller with a targeted cell recovery of 5,000 cells. 3′ mRNA-seq gene expression libraries were prepared per the Chromium Single Cell 3' User Guide (10x Genomics, CG000204). Library quality was verified using TapeStation 4200 (Agilent) and libraries were quantified by Qubit fluorometer (Thermo Fisher) prior to pooling. Library pools were sequenced at the National Jewish Health Genomics Facility on an Illumina NEXTSEQ 2000, 28x10x10x90 bp, paired-end sequencing.