|
| Status |
Public on May 26, 2011 |
| Title |
SiTIF+ rep2 |
| Sample type |
RNA |
| |
|
| Source name |
HMEC-TR, SiRNA anti-TIF1γ, 5 ng/ml TGFβ1 for 24h
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: Human mammary epithelial cells infected with retrovirus carrying hTERT and the oncogenic allele of H-Ras, H-RasV12 described in (Elenbaas B, Spirio L, Koerner F, Fleming MD, Zimonjic DB, Donaher JL, Popescu NC, Hahn WC, and Weinberg RA (2001) Human breast cancer cells generated by oncogenic transformation of primary mammary epithelial cells. Genes Dev 15: 50-65)
|
| Treatment protocol |
We performed transcriptomic microarrays analysis on control HMEC-TR cells (transfected by scramble si-RNA) and cells inactivated for TIF1γ or Smad4 (SiRNA strategy) and treated or not with TGFβ1 for 24 hours. Inactivation of TIF1γ and Smad4 expression was checked by Western Blot.
|
| Growth protocol |
Human mammary epithelial cells (HMEC-TR) kindly provided by RA Weinberg have been previously described (Elenbaas B. et al. 2001 and Morel A.P. et al. 2008). These cells were cultured in 1:1 Dulbecco’s Modified Eagle’s Medium DMEM/HAMF12 medium (Invitrogen) complemented with 10% FBS (Cambrex) and 100 U/ml penicillin-streptomycin (Invitrogen), 2 mM L glutamine (Invitrogen), 10 ng/ml human epidermal growth factor (hEGF, PromoCell), 0.5 mg/ml hydrocortisone (Sigma) and 10 mg/ml insulin (Sigma).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from the transfected HMEC-TR cells was isolated by Trizol and miRNeasy Mini extraction kit (Qiagen). RNA quality was assessed by an Agilent Bioanalyzer (model 2100; Agilent Technologies).
|
| Label |
biotin
|
| Label protocol |
As per manufacturer (Affymetrix)
|
| |
|
| Hybridization protocol |
As per manufacturer (Affymetrix)
|
| Scan protocol |
The Arrays were scanned using the Gene Chip Scanner 3000. Then CEL files were generated using the Affymetrix GeneChip Operating Software (GCOS)
|
| Description |
rimo10
|
| Data processing |
CHP files were processed using MAS 5.0 analysis algorithm with Affymetrix GeneChip Operating Software (GCOS)
|
| |
|
| Submission date |
Apr 07, 2011 |
| Last update date |
May 26, 2011 |
| Contact name |
Cedric Hesling |
| E-mail(s) |
hesling@lyon.fnclcc.fr
|
| Organization name |
CRCL
|
| Lab |
Equipe Ruth Rimokh
|
| Street address |
24 rue Laënnec
|
| City |
Lyon |
| ZIP/Postal code |
69008 |
| Country |
France |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE28448 |
Antagonistic regulation of EMT by TIF1γ and Smad4 in mammary epithelial cells |
|
