2024-12-06
ENA first public
ERC000011
ENA-CHECKLIST
Genentech, Computational Sciences
INSDC center name
public
INSDC status
ERS21395703
SRA accession
E-MTAB-14584:Sample 3
Submitter Id
ArrayExpress
broker name
T cell
cell_type
not collected
collection date
house mouse
common name
Protocols: CD8+ T cells were purified from spleens of OT-I mice by negative selection with magnetic beads (EasySep, Stemcell Technologies). After purification, cells were >95% CD8+ T cells. In each well of a 96-well plate, 0.5x106 of the purified CD8+ T cells/ml were cultured in complete media (RPMI 1640, 10% FBS (Gibco), 1% 2 mM L-glutamine (Life Technologies), 1% HEPES (Life Technologies), 1 nM Sodium Pyruvate (Life Technologies), 1% non-essential amino acids (Life Technologies), 100 U/ml penicillin (Gibco) and 100 μg/ml streptomycin-sulfate (Gibco), 0.05 mM 2-mercaptoethanol (Sigma) with recombinant human IL-2 (10 ng/ml, Roche) with or without 10 ng/ml OVA(257–264) (SIINFEKL) peptide in the presence or absence of 25 ng/ml murine IL-27 (R&D Systems). For repeated peptide stimulation, 10 ng/ml OVA(257–264) peptide was added daily for 4–6 days. Cells were checked daily, passaged, and cultured with fresh complete media containing cytokines as required. After day 5 cells were washed and stained for flow cytometry. ATAC-seq libraries were generated from fresh cells using the ATAC-Seq Kit (Active Motif, catalog# 53150).  Libraries were quantified using the Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific) and profiled using the D5000 ScreenTape on TapeStation 4200 (Agilent Technologies).
description
adult
developmental stage
not collected
geographic location (country and/or sea)
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
spleen
organism part
Mus musculus
scientific_name
C57BL/6J
strain
Sample 3
title