2025-12-13
ENA first public
ERC000011
ENA-CHECKLIST
VIB-UGent Inflammation Research Center
INSDC center name
public
INSDC status
ERS27232432
SRA accession
E-MTAB-16099:Exp2_WT_lung_PBS
Submitter Id
ArrayExpress
broker name
immune and non-immune
cell type
not collected
collection date
house mouse
common name
Protocols: Exp2: Mice were treated with PBS, 1 x 100 ng IL-33 intratreacheally or 3 x 100 ng IL-33 intratracheally. Mice were sacrificed 24h after the last treatment and single cell lung suspensions were created. Cells were incubated with FACS antibodies as well as TotalSeq A hashtag antibodies. 12000 CD45-, 3000 CD45+ CD11c+ SiglecF+ alveolar macrophages, 3000 CD45+ lineage- CD11c- SiglecF- CD90+ ST2+ ILC2s and 12000 AM and ILC2 excluded CD45+ cells were FACS purified per mouse and pooled together per condition. Cells were diluted to a final concentration of 1000/μl and loaded on the 10X Chromium device. The DNA libraries were prepared using the GemCode Single Cell 3′ Gel Bead and Library kit, version NextGEM 3.1 (10× Genomics) according to the manufacturer's instructions with the addition of amplification primers (3 nM, 5′CCTTGGCACCCGAGAATT*C*C and 5′GTGACTGGAGTTCAGACGTGTGC*T*C) during cDNA amplification to enrich the TotalSeq-A cell surface protein and/or hashtag oligos. Size selection with SPRIselect Reagent Kit (Beckman Coulter, B23318) was used to separate amplified cDNA molecules for 3′ gene expression and cell surface protein construction. TotalSeq-A protein library construction including sample index Polymerase Chain Reaction (PCR) using Illumina's Truseq Small RNA primer sets and SPRIselect size selection was performed according to the manufacturer's instructions.
description
PBS treated
disease
wild type genotype
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
lung
organism part
Mus musculus
scientific_name
female
sex
C57BL6J
strain
Exp2_WT_lung_PBS
title