2024-06-29
ENA first public
ERC000011
ENA-CHECKLIST
SAMEA113825297
External Id
Laboratory for Molecular Infection Medicine Sweden Umeå university Department of molecular biology
INSDC center name
2023-06-19T16:24:01Z
INSDC last update
public
INSDC status
ERS15819466
SRA accession
E-MTAB-13089:rep1_hto
Submitter Id
8 to 12
week
age
ArrayExpress
broker name
CD4-positive helper T cell
cell_type
not collected
collection date
house mouse
common name
Protocols: CD4cre+HDAC1f/f mice were crossed to IL-13 dtTomato reporter mice. Between 8 to 10 weeks old CD4cre-HDAC1f/f dtTomato+ and CD4cre+HDAC1f/f dtTomato+ mice on C57BL/6 background were sensitized by intratracheal injection on day 0 and day 5 with 10 µg House dust mite (HDM) extract, challenged intranasally on day 12 and 14 with 25 µg of HDM, and sacrificed on day 16. After bronchoalveolar lavage, lungs were perfused with PBS, homogenized, digested and IL-13+, IL-13- effector (CD44hiCD62L-) and naive (CD44loCD62L+) CD4+ T cells were sorted by FACS. As control, IL-13- effector and naive cells were sorted from untreated CD4cre-HDAC1f/f dtTomato+ and CD4cre+HDAC1f/f dtTomato+ mice. Sorted cells from WT and KO mice were incubated with different DNA-barcoded antibodies for sample demultiplexing. Nucleic acids extracted as an integrated part of the single cell library construction protocol. All single-cell suspensions were counted and diluted according to manufacturer recommendations before loading onto a Chromium Controller (10X Genomics, Pleasanton, CA, USA). Single-cell RNA-seq libraries were generated using the Next GEM Single Cell 3’ Reagent Kit (v3.1, 10x Genomics, Pleasanton, CA, USA) according to the manufacturer’s instructions aiming for a maximum cell recovery of 10,000 cells.
description
adult
developmental stage
mixed CD4cre-HDAC1f/f dtTomato+ and CD4cre+HDAC1f/f dtTomato+
genotype
not collected
geographic location (country and/or sea)
mixed
individual
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
lung
organism part
feature barcoding
readout
Mus musculus
scientific_name
female
sex
C57BL/6
strain
rep1_hto
title