|
| Status |
Public on May 09, 2016 |
| Title |
resting NK cells_normoxia_biological rep4 |
| Sample type |
RNA |
| |
|
| Source name |
resting NK cells_normoxia
|
| Organism |
Homo sapiens |
| Characteristics |
subject status: healthy donor
tissue: peripheral blood
cell type: Natural killer (NK) cells
cultured under: Normoxia for 22hrs
treated with: none (control)
|
| Treatment protocol |
Parallel NK cell cultures were grown under normoxia or hypoxia for 22 hours with or without 45 ng/mL IL-15 present during the final 6 hours.
|
| Growth protocol |
Freshly isolated NK cells were plated at one million cells per mL RPMI 1640 supplemented with 10% fetal bovine serum and 2 mM L-glutamine in a 3ºC humidified atmosphere of 5% carbon dioxide (normoxia). One percent oxygen (hypoxia) was established in an oxygen controlled carbon dioxide incubator with a nitrogen gas line.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was prepared from NK cells stored in RNAlater using a spin column protocol followed by DNAse treatment according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol.
|
| |
|
| Hybridization protocol |
Hybridization (16h x 45°C) was processed according to the standard Affymetrix protocol.
|
| Scan protocol |
Affymetrix GeneArray Scanner3000
|
| Description |
N_ctrl_P22
|
| Data processing |
The data were analyzed with a commercial software called JMP Genomics, version 6, from SAS. Gene expression profiling was performed using arrays of HuGene-2_0-st-type from Affymetrix. A Custom CDF Version 17 with Entrez based gene definitions was used to annotate the arrays. The Raw fluorescence intensity values were normalized applying quantile normalization, RMA background correction and Medianpolish Probeset Summary
|
| |
|
| Submission date |
Jun 24, 2015 |
| Last update date |
May 09, 2016 |
| Contact name |
Carsten Sticht |
| Organization name |
University Heidelberg
|
| Department |
ZMF
|
| Street address |
Theodor-Kutzer-Ufer
|
| City |
Mannheim |
| ZIP/Postal code |
68169 |
| Country |
Germany |
| |
|
| Platform ID |
GPL17930 |
| Series (1) |
| GSE70214 |
Short-term hypoxia synergizes with interleukin 15 priming in driving glycolytic gene transcription and supports human natural killer cell activities. |
|
