|
| Status |
Public on May 20, 2019 |
| Title |
Notch2 WT MSCs R3 |
| Sample type |
RNA |
| |
|
| Source name |
bone marrow derived MSCs
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57B6
genotype: Notch2 WT
original sample name: R3
|
| Treatment protocol |
Cells were incubated overnight in a 1:1 mixture of medium (DMEM w/o FCS) and PBS containing 1 μM Tat-Cre protein (HTNC). BMSCs were then cultured at least 8 days after transduction in DMEM supplemented with PS and 10% FCS before co-culture experiments were performed. Primary CLL cells were cultured on MSCs for 5 days, then washed off and remaining CLL cells removed with anti-CD19 beads.
|
| Growth protocol |
PBMCs from C57B6 Notch2fl/fl mice were generated from the bone marrow. Adherend cells were cultured in MEM Alapha + GlutaMAX medium (Invitrogen) supplemented with 10% fetal calf serum (PAA, Cölbe, Germany), 10% horse Serum.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted by using the RNeasy Mini Kit (Qiagen). Total-RNA concentration and purity of total-RNA samples was measured with the NanoDrop 1000 spectrophotometer (NanoDrop Technologies/Thermo Scientific).
|
| Label |
Biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the Affymetrix ExpressKit protocol starting from 200 ng total RNA.
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45°C on GeneChip MG-430_2.0 arrays. GeneChips were washed and stained in the Affymetrix Fluidics Station 450 using the Affymetrix Hybridization, Wash and Stain Kit.
|
| Scan protocol |
Arrays were scanned in a GeneChip 3000 scanner with G7 update.
|
| Data processing |
The images were analyzed with RMA as implemented in PartekGS.
|
| |
|
| Submission date |
Jun 02, 2017 |
| Last update date |
May 20, 2019 |
| Contact name |
Ludger Klein-Hitpass |
| E-mail(s) |
ludger.klein-hitpass@uni-essen.de
|
| Phone |
+49 201 723 85552
|
| Organization name |
Institut fuer Zellbiologie
|
| Department |
Universitaetsklinikum
|
| Lab |
BioChip Lab
|
| Street address |
Virchowstr. 173
|
| City |
Essen |
| ZIP/Postal code |
D-45122 |
| Country |
Germany |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE99614 |
Notch2 in mesenchymal stromal cells regulates canonical Wnt signalling in CLL cells |
|
