|
| Status |
Public on Dec 31, 2015 |
| Title |
Fibroblast, apoptotic bodies (24h), biological rep1 |
| Sample type |
RNA |
| |
|
| Source name |
apoptotic bodies collected 24h after CHX treatment, fibroblasts
|
| Organism |
Homo sapiens |
| Characteristics |
cell source: primary dermal fibroblasts
cell type: apoptotic bodies
|
| Treatment protocol |
To induce apoptosis, the cells were incubated with cycloheximide (5µg/ml) in hypoxia (2% O2) for 24 hours. Afterwards, the apoptotic bodies were collected.
|
| Growth protocol |
Cells were cultured in endothelial growth medium (PromoCell) to confluency before starting apoptosis.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from apoptotic bodies with the ZR RNA MicroPrep kit (Zymo Research) according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Labelling was prepared according to the standard FlashTag (Genisphere) protocol.
|
| |
|
| Hybridization protocol |
Hybridization was performed for 16 hr at 48°C and 60 rpm on GeneChip miRNA 3.0 Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450 using the Fluidics script FS450-0002.
|
| Scan protocol |
GeneChip miRNA 3.0 Arrays were scanned on a GeneChip Scanner 3000 7G (Affymetrix).
|
| Description |
miRNA expression data from fibroblast apoptotic bodies.
|
| Data processing |
The image data were analyzed with the miRNA QC Tool software for quality control. The expression values were summarized and normalized with robust multi-array average (RMA) using RMA package in Bioconductor 2.5 under R2.10. Only miRNAs that were detectable in all three arrays for each group and whose expression levels were found to have changed significantly (P<0.05), at least by twofold, were considered to be differentially expressed.
|
| |
|
| Submission date |
Jun 21, 2013 |
| Last update date |
Dec 31, 2015 |
| Contact name |
Bernd Denecke |
| E-mail(s) |
bernd.denecke@rwth-aachen.de
|
| Phone |
+49 241 8089918
|
| Organization name |
RWTH Aachen University
|
| Department |
IZKF Aachen
|
| Street address |
Pauwelsstrasse 30
|
| City |
Aachen |
| State/province |
NRW |
| ZIP/Postal code |
52074 |
| Country |
Germany |
| |
|
| Platform ID |
GPL16384 |
| Series (1) |
| GSE48218 |
Novel critical insights into the mechanism of cell-based therapy after myocardial infarction |
|
