title: GSM8210142: RNA_MatN_KEP_aIL1B_1; Mus musculus; RNA-Seq
library_construction_protocol: LT-HSCs, MPPs, CMPs and GMPs were enriched by neutrophil depletion. Bone marrow single cell suspensions were incubated with 10μg/ml anti-Ly6G (1A8, BioXcell) before neutrophil depletion using sheep anti-rat magnetic beads (Dynabeads, Life Technologies) and subsequently stained with cell surface markers. For sorting pre-neutrophils and neutrophils from bone marrow, blood and lung, cells were first incubated with 500μg/ml purified mouse IgG+10% FCS (BM) or rat anti-mouse CD16/CD32 antibody (blood and lung), stained with cell surface markers and sorted using gating strategies for HSPC populations or CD45+ CD11b+ Ly6G+ 7-AAD- markers for sequencing of blood and lung neutrophils. RNA was isolated following RNeasy Mini Kit (Qiagen) protocol, using 80% ethanol instead of RPE buffer. Smart-seq2 library preparation was performed according to the Smart-seq2 protocol, using 2100 Bioanalyzer System (Agilent) for quality control.