|
| Status |
Public on Aug 02, 2011 |
| Title |
shZBP-89, biological rep2 |
| Sample type |
RNA |
| |
|
| Source name |
erythroblast transduced with shZBP-89
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: Erythroid progenitors on day 6 of differentiation
genotype/variation: transduced with lentivirus expressing short hairpins against ZBP-89
|
| Treatment protocol |
Expanding hCD34+ cells were seeded in fresh medium 24 hrs post-transduction, and cultured for an additional 24 hrs prior to the start of selection with puromycin. The empty pLKO.1ps vector and shZBP-89 in the same vector backbone are used.
|
| Growth protocol |
De-identified cryogenically preserved primary human CD34+ (hCD34+) cells were obtained from the Yale Center of Excellence in Molecular Hematology (New Haven, Connecticut). They were prepared by magnetically sorting mononuclear samples of G-CSF mobilized peripheral blood from donors. For erythroid ex vivo differentiation, hCD34+ progenitor stem cells were expanded for 6 days in StemSpan SFEM Medium with 1X CC100 cytokine mix (StemCell Technologies Inc.) and 2% P/S. On day 6 of expansion, cells were reseeded into erythroid differentiation medium (StemSpan SFEM Medium containing 2% P/S, 20 ng/ml SCF, 1 U/ml Epo, 5 ng/ml IL-3, 2 uM dexamethasone, and 1 uM β-estradiol)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Extraction of total RNA was performed using Qiagen RNeasy Plus Mini Kit according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
According to standard Affymetrix protocols.
|
| |
|
| Hybridization protocol |
According to standard Affymetrix protocols.
|
| Scan protocol |
According to standard Affymetrix protocols.
|
| Data processing |
dCHIP program using Affymetrix default analysis settings and global scaling as normalization method
|
| |
|
| Submission date |
Aug 01, 2011 |
| Last update date |
Aug 02, 2011 |
| Contact name |
Andrew Jonghan Woo |
| E-mail(s) |
andrew.woo@childrens.harvard.edu
|
| Phone |
6179192028
|
| Fax |
6177300222
|
| Organization name |
Children's Hospital Boston / Harvard Medical School
|
| Department |
Hematology / Oncology
|
| Lab |
Alan B Cantor Lab
|
| Street address |
1 Blackfan Circle, Karp Res Bldg
|
| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02115 |
| Country |
USA |
| |
|
| Platform ID |
GPL571 |
| Series (1) |
| GSE31092 |
Expression analysis of ZBP-89 deficient human primary erythroid progenitors |
|
