|
| Status |
Public on Aug 29, 2012 |
| Title |
Stat5b prolactin rep3 |
| Sample type |
RNA |
| |
|
| Source name |
MCF7 human breast cancer cell line expressing exogenous Stat5b, treated with 10nM human prolactin for 4 hours
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: MCF7 human breast cancer
exogenous protein expression: Stat5b
hormone treatment: prolactin
|
| Treatment protocol |
MCF7 cells were infected with adenovirus expressing Stat5a or Stat5b for 90 min in serum free media and cultured for an additional 24 hr in regular growth media. Cells were serum-starved 16 hr prior to treatment with prolactin for 4 hr in serum-free media.
|
| Growth protocol |
MCF7 human breast cancer cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum, 2mM L-glutamine, and 50 IU/ml penicillin-50ug/ml streptomycin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was harvested using the Qiagen RNeasy Kit following the recommended protocol.
|
| Label |
biotin
|
| Label protocol |
Amplified single-stranded cDNA was fragmented and labeled using the Encore™ cDNA Biotin Module. The enzymatically and chemically fragmented product (50-100 nt) was labeled via the attachment of biotinylated nucleotides onto the 3'-end of the fragmented cDNA.
|
| |
|
| Hybridization protocol |
The resultant fragmented and labeled cDNA was added to the hybridization cocktail in accordance with the NuGEN™ guidelines for hybridization onto Affymetrix GeneChip® arrays. Hybridization was run for 16-18 hours at 45°C in an Affymetrix GeneChip® Hybridization Oven 640. Following hybridization, the array was washed and stained on the GeneChip® Fluidics Station 450.
|
| Scan protocol |
Arrays were inserted into the Affymetrix autoloader carousel and scanned using the GeneChip® Scanner 3000
|
| Data processing |
Raw intensity files from the Affymetrix Human Gene 1.0 ST GeneChip arrays were processed using Affymetrix Expression Console version 1.1. Gene-level expression measurements were computed using the iterPLIER algorithm on the “core” probesets and exported with annotation release 30, dated November 9th, 2009.
|
| |
|
| Submission date |
May 04, 2012 |
| Last update date |
Aug 31, 2012 |
| Contact name |
Adam Ertel |
| Organization name |
Thomas Jefferson University
|
| Department |
Cancer Biology
|
| Lab |
BLSB 1009
|
| Street address |
233 S 10TH ST
|
| City |
PHILADELPHIA |
| State/province |
PA |
| ZIP/Postal code |
19107 |
| Country |
USA |
| |
|
| Platform ID |
GPL6244 |
| Series (1) |
| GSE37781 |
Prolactin-inducible Stat5a-mediated and Stat5b-mediated gene profiles |
|
