|
| Status |
Public on Sep 09, 2012 |
| Title |
HCLS1 shRNA |
| Sample type |
RNA |
| |
|
| Source name |
Bone marrow CD34+ cells, HCLS1 shRNA
|
| Organism |
Homo sapiens |
| Characteristics |
disease state: healthy
tissue: bone marrow
cell type: CD34+ cells
treatment: HCLS1 shRNA
|
| Treatment protocol |
CD34+ hematopoietic cells were transduced with lenitvirus-based RFP-expressed shRNA constructs targeting HCLS1 mRNA (two different shRNA constructs were used simultaneously) or scrambled control shRNA. On day 4 after transduction, transduced RFP+ cells were sorted and total RNA was isolated.
|
| Growth protocol |
Cells were cultured in ex-vivo medium supplemented with 10% HSA, 20 ng/ml IL-3, 20 ng/ml IL-6, 20 ng/ml TPO, 50 ng/ml SCF and 50 ng/ml Flt-3L.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Extraction of total RNA was performed using the RNeasy Micro Kit (Qiagen) according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
The GeneChip WT cDNA Synthesis and Amplification Kit was used to make double-stranded cDNA from total RNA, which was then labeled with biotin (GeneChip WT Terminal Labeling Kit).
|
| |
|
| Hybridization protocol |
After chemical fragmentation of the biotin-labeled cRNA targets, they were hybridized to Affymetrix Human Exome microarrays using the Fluidics Station 450.
|
| Scan protocol |
Microarrays were scanned using the Affymetrix Genechip Scanner 3000 with Genechip Operating Software 1.4 (Affymetrix, Santa Clara, CA).
|
| Description |
shRNA_HS1
Gene expression data from CD34+ cells transduced with HCLS1 shRNA.
|
| Data processing |
The data were analyzed with Affymetrix Expression Console software with RMA-sketch exon-level analysis settings.
|
| |
|
| Submission date |
Sep 09, 2012 |
| Last update date |
Sep 09, 2012 |
| Contact name |
Siarhei Kandabarau |
| E-mail(s) |
kandabarau.siarhei@mh-hannover.de
|
| Organization name |
Hannover Medical School
|
| Street address |
Carl-Neuberg-Str 1
|
| City |
Hannover |
| ZIP/Postal code |
30625 |
| Country |
Germany |
| |
|
| Platform ID |
GPL10739 |
| Series (1) |
| GSE40712 |
Expression data from CD34+ hematopoietic cells transduced with control or anti-HCLS1 shRNA |
|
