|
Status |
Public on Mar 29, 2020 |
Title |
Tcm_PEneg_B |
Sample type |
RNA |
|
|
Source name |
primary T lymphocytes derived from human peripheral blood mononuclear cells (PBMC)
|
Organism |
Homo sapiens |
Characteristics |
cell type: effector memory T lymphocytes culture condition: Effector memory T lymphocytes were stimulated with PMA/Ionomycin, PE-labelled within a GM-CSF Cytokine Secretion Assay (Miltenyi Biotec) and sorted for PEpos/neg before total RNA extraction label: PEneg
|
Treatment protocol |
Tem were stimulated for 3h with PMA/ionomycin and labelled for GM-CSF secretion with a GM-CSF cytokine secretion kit (Miltenyi Biotec) before sorting for PE+/- cells with a BD Sorter Aria II
|
Growth protocol |
Effector memory T lymphocytes were isolated by magnetic enrichment for CD4 (Miltentyi Biotec) and sorted for CD4+CD25-CD45RA-CCR7- and incubated overnight in complete media
|
Extracted molecule |
total RNA |
Extraction protocol |
PE+/- cells were resuspended in Trizol. Total RNA was extracted with Direct-Zol RNA mini Prep (Zymo Research)
|
Label |
biotin
|
Label protocol |
100 ng of total RNA were labelled with the nCounter® Human miRNA Expression Assay codeset according to the manufacturers protocol
|
|
|
Hybridization protocol |
Samples were hybridized overnight at 65C
|
Scan protocol |
Samples were run on a Nanostring Sprint Profiler
|
Description |
Human peripheral blood mononuclear cells were separated from peripheral blood using Ficoll-Paque Plus (GE Healthcare). CD4+ T lymphocytes were further isolated by positive selection using magnetic microbeads (Miltenyi Biotec). Effector Memory T cell subsets were then sorted based on the expression of the following surface markers: CD4+CD25–CD45RA-CCR7-. nCounter® Human miRNA Expression Assay
|
Data processing |
Raw data was processed with nSolver 3.0 Software (Nanostring). Reads were normalized to internal postive & negative controls and to the 25 highest expressed miRNAs. Genes were considered expressed if at least one sample had a read >125 and maximum 1 sample with a read <27
|
|
|
Submission date |
Nov 26, 2018 |
Last update date |
Mar 29, 2020 |
Contact name |
Chiara Balestrieri |
E-mail(s) |
balestrieri.c@gmail.com
|
Organization name |
IRCCS San Raffaele Scientific Institute
|
Department |
Center for Omics Sciences
|
Street address |
Via Olgettina 58
|
City |
Milan |
ZIP/Postal code |
20132 |
Country |
Italy |
|
|
Platform ID |
GPL25243 |
Series (2) |
GSE122943 |
A molecular network regulating the pro-inflammatory phenotype of human memory T lymphocytes (Nanostring II) |
GSE122946 |
A molecular network regulating the proinflammatory phenotype of human memory T lymphocytes |
|