|
Status |
Public on Nov 26, 2019 |
Title |
SLE_CD8_1 |
Sample type |
genomic |
|
|
Source name |
CD8+ T cell DNA
|
Organism |
Homo sapiens |
Characteristics |
cell type: CD8+ T cell disease status: lupus
|
Extracted molecule |
genomic DNA |
Extraction protocol |
Peripheral blood mononuclear cells (PBMCs) were extracted from whole blood via Ficoll-Paque PLUS density centrifugation (GE Healthcare Bio-Sciences). Total T cells were isolated using negative selection by magnetic bead separation with the Human Pan T cell Isolation II kit (Miltenyi Biotec). Flow cytometry sorting was used to isolate CD3+CD56-TCRab+CD8+ T cells using the following fluorophore-conjugated antibodies: APC anti-human CD56 (clone: HCD56), FITC anti-human CD8a (clone: RPA-T8), Pacific Blue anti-human CD3 (clone: HIT3a), and PE anti-human alpha/beta-TCR (clone: IP26) (BioLegend). DNA was extracted from CD8+ T cells using the DNeasy Blood and Tissue kit (Qiagen) and bisulfite converted with the EZ DNA Methylation kit (Zymo Research) for DNA methylation analysis.
|
Label |
biotin
|
Label protocol |
standard Illumina protocol
|
|
|
Hybridization protocol |
standard Illumina protocol
|
Scan protocol |
standard Illumina protocol
|
Description |
lupus CD8+ T cell
|
Data processing |
Methylation beta values were calculated from raw intensity values using GenomeStudio Methylation module software (Illumina; ver 1.9.0) after subtracting background, normalization to control probes, error correction using the Illumina custom model, and correction for false discovery rate.
|
|
|
Submission date |
Nov 27, 2018 |
Last update date |
Nov 27, 2019 |
Contact name |
Amr Sawalha |
E-mail(s) |
asawalha@pitt.edu
|
Organization name |
University of Pittsburgh
|
Street address |
4401 Penn Avenue
|
City |
Pittsburgh |
State/province |
PA |
ZIP/Postal code |
15224 |
Country |
USA |
|
|
Platform ID |
GPL23976 |
Series (1) |
GSE123003 |
Hypomethylation of STAT1 and HLA-DRB1 is associated with type-I interferon-dependent HLA-DRB1 expression in lupus CD8+ T cells |
|