characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-10-31T00:26:54Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-10-31T00:26:54Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA113400781
characteristics.characteristic: External Id
characteristics.text: UNIVERSITY OF MANCHESTER
characteristics.characteristic: INSDC center name
characteristics.text: 2023-10-31T00:26:54Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-10-31T00:26:54Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS15395922
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12864:WT
characteristics.characteristic: Submitter Id
characteristics.text: 12
characteristics.unit: week
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: innate lymphoid cell
characteristics.characteristic: cell_type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: We performed single-cell RNA sequencing from small intestine ILC as previously described (Fiancette et al., 2021). Briefly, Id2-CreERT2 ROSA26-tdRFP Bcl6-fl/fl and Id2-CreERT2 ROSA26-tdRFP control mice were treated with 5mg of tamoxifen in corn oil 5 times over the course of 2 weeks via oral gavage. Mice were sacrificed through cervical dislocation 3 weeks after the last dose. Fat and Peyer’s patches were removed from the small intestine, which was opened longitudinally and washed vigorously in PBS. Tissue was incubated for two concurrent rounds in PBS containing 1 mM EDTA, 1 mM dithiothreitol and 5 % FBS for 25 min at 37 °C with constant mixing. Then, tissues were washed in PBS and digested in complete RPMI with in 1 mg/ml collagenase VIII (Sigma-Aldrich) and 20 μg/ml DNase (Sigma-Aldrich) for 25 min at 37 °C. The digested tissues were filtered through a 70 μm nylon strainer and stained for fluorescence-activated cell sorting. Live CD45+ B220- CD11c- CD11b- CD3- CD5- TCRβ- TCRγδ- CD90.2+ CD127+ ROSA26-tdRFP+ ILC were sorted into PBS 10% FBS using a BD FACS Aria II. Sorted cells were passed through the 10x Chromium (10x Genomics) system as indicated on the manufacturer’s protocol (CG000315 Rev B), at a target recovery rate of 10,000 cells. cDNA library generation was performed as indicated in the protocol for Chromium Single Cell 3’ Reagent kits v3.1 (10x Genomics). Resulting libraries were prepared in parallel in a single batch.
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.characteristic: disease
characteristics.text: Id2-CreERT2 ROSA26-tdRFP
characteristics.characteristic: genotype
characteristics.text: pool of 2 individuals
characteristics.characteristic: individual
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: small intestine lamina propria
characteristics.characteristic: organism part
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: male
characteristics.characteristic: sex
characteristics.text: C57BL/6J
characteristics.characteristic: strain
characteristics.text: WT
characteristics.characteristic: title