2025-06-27
ENA first public
ERC000011
ENA-CHECKLIST
BioISI - Faculty of Sciences, University of Lisbon
INSDC center name
public
INSDC status
ERS23841414
SRA accession
D1AScr
Submitter Id
Peripheral blood mononuclear cells (PBMC) were isolated from buffy coats by density gradient centrifugation using Lymphosep (Biowest), followed by isolation of naive CD4+T cells by negative selection using the MojoSort� Human CD4 Na�ve T Cell Isolation Kit (BioLegend). Naive CD4+ T cells were stimulated for 40 hours with CD3/CD28 dynabeads (Gibco) (1bead:10 cells) + 10U/mL of hrIL-2. After 40 hours, stimulation beads were removed, cells were washed with phosphate buffered saline (PBS) and resuspended in serum-free RPMI to reach a final concentration of 2 million cells/mL/well and plated on a 24-well plate. Control or miR-34c-5p-specific-antagomiRs were directly added to the plated cells to reach a final concentration of 1�M. AntagomiR-treated cells were incubated for 2 hours. After treatment, the medium was replaced, and cells were cultured in RPMI-1640 + GlutaMAX (Gibco) medium supplemented with 10% FBS, 1% nonessential amino acids, 1%, sodium pyruvate, 1% penicillin/streptomycin, 0.1% gentamycin and 10U/mL of hrIL-2for 100 hours more. Total RNA was extracted with miRNeasy Mini Kit (Qiagen).
Treatment
Naive CD4+ T cells
cell_type
2022-08
collection date
human
common name
scramble-antagomiR-treated in vitro stimulated naive CD4+ T cells from donor 1
description
Portugal
geographic location (country and/or sea)
Homo sapiens
organism
Homo sapiens
scientific_name
D1AScr
title