organism: Mus musculus
description: Protocols: Mesenteric lymph nodes were collected in PBS/BSA/EDTA and passed through a 70 μm filter. Single cell suspensions were washed in PBS/BSA/EDTA and stained with fluorochrome-conjugated antibodies in PBS/BSA/EDTA for 20 min at 4 oC in the dark. Cells were washed and resuspended in PBS/BSA/EDTA/DNase1 for FACS sorting. Foxp3+ Tregs were sorted as live CD45+Lineage-CD4+TCRβ+Foxp3-YFP+ cells. Lineage included CD8, TCRγδ, B220, CD11b, CD11c, Gr1. Dead cells were excluded using DAPI. For gene expression analysis by RNA sequencing, small bulks of 300 Treg cells were sorted into 2μL lysis buffer (0.8% Triton X-100 and 2 U/μL RNase inhibitor) in a 96-well plate Sequencing libraries were prepared following the Smart-seq2 protocol (Picelli et al., 2013) with own in-house indexing primers (Lamble et al., 2013)
identifiers.namespace: BioSample
identifiers.id: SAMEA111415843
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2023-01-01
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2023-01-01
attributes.tag: External Id
attributes.value: SAMEA111415843
attributes.tag: INSDC center alias
attributes.value: OXFORD
attributes.tag: INSDC center name
attributes.value: University of Oxford
attributes.tag: INSDC first public
attributes.value: 2023-01-01T00:28:50Z
attributes.tag: INSDC last update
attributes.value: 2023-01-01T00:28:50Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-12226:mLN6
attributes.tag: age
attributes.value: 11 to 14
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_type
attributes.value: regulatory T cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: DGAT1-/-
attributes.tag: individual
attributes.value: 6
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: mesenteric lymph node
attributes.tag: replicate
attributes.value: 6
attributes.tag: sample name
attributes.value: E-MTAB-12226:mLN6
attributes.tag: sex
attributes.value: mixed
attributes.tag: strain
attributes.value: C57BL/6