title: Sample 3
organism: Mus musculus
description: Protocols: Spleen and lymph nodes (inguinal and axillary) were harvested from mice under sterile conditions, mechanically dissociated into single cell suspension and red blood cells were lysed. Cells were then sorted by negative selection following the manufacturer's instructions. After sorting, cells were resuspended at 1x106 cells/mL in complete RPMI media (10% FBS, 1% NaPy, 1% Glutamine, 1% Pen/Strep, 0,1% b-mercaptoethanol) supplemented with 100U/mL IL2. For OT1 mice, media was further complemented with 2 mg/mL of SIINFELK peptide, while for C57J/B6 mice polyclonal activation was achieved using 5 mg/mL plated-bound aCD3 and 0,5 mg/mL soluble aCD28. Cells were further treated with BSA, linoleic acid, oleic acid or arachidonic acid. OT1 cells were sorted with >95% purity as previously described, activated for 48 hours in the presence or absence of LA, counted and resuspended in RLT lysis buffer (Qiagen). RNA was isolated from purified cells using RNeasy Mini kits, and RNA concentrations were determined using Nanodrop. RNA-seq, Random sequencing of whole transcriptome.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2023-06-01
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2023-06-01
attributes.tag: External Id
attributes.value: SAMEA112770668
attributes.tag: INSDC center alias
attributes.value: Microbiome and Antitumor Immunity Dpt. of Experimental Oncology European Institute of Oncology, Milan, Italy
attributes.tag: INSDC center name
attributes.value: Microbiome and Antitumor Immunity Dpt. of Experimental Oncology European Institute of Oncology, Milan, Italy
attributes.tag: INSDC first public
attributes.value: 2023-06-01T00:15:37Z
attributes.tag: INSDC last update
attributes.value: 2023-06-01T00:15:37Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-12645:Sample 3
attributes.tag: age
attributes.value: 2
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_type
attributes.value: mononuclear cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: mixed spleen and lymph node
attributes.tag: sample name
attributes.value: E-MTAB-12645:Sample 3
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: male
attributes.tag: stimulus
attributes.value: 5 mg/mL plated-bound aCD3 and 0,5 mg/mL soluble aCD28
attributes.tag: strain
attributes.value: C57BL/6J