|
| Status |
Public on Apr 10, 2020 |
| Title |
siCAV1+TGF-β rep 2 |
| Sample type |
RNA |
| |
|
| Source name |
primary hepatocytes treated with Cav1 siRNA and TGFBeta
|
| Organism |
Mus musculus |
| Characteristics |
sirna: Caveolin-1
treatment: TGFBeta
|
| Treatment protocol |
After attachment, primary hepatocytes were transfected overnight with 10 nM siRNA targeting CAV1 or a negative control siRNA using RNAiMAX transfection reagent according to the manufacturer’s instructions. Next day, the hepatocytes were treated with/without 5 ng/ml recombinant TGF-β1 and cultured additional 48 h in starvation medium.
|
| Growth protocol |
Primary hepatocytes were prepared from 12 week old C57BL/6 mice via the two-step collagenase perfusion method.Then, isolated hepatocytes were plated on 6-well plates and cultured in Williams E medium with 1% l-glutamine, 1% Penicillin/Streptomycin, 0.1% dexamethasone and 10% fetal bovine serum. After hepatocyte attachment (~4 h), culture medium was changed to starvation medium (no dexamethasone and fetal bovine serum).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using InviTrap Spin Universal RNA Mini Kit from all sample groups according to the manufacturer’s instruction.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol.
|
| |
|
| Hybridization protocol |
Hybridization (16h x 45°C) was processed according to the standard Affymetrix protocol.
|
| Scan protocol |
Affymetrix GeneArray Scanner3000
|
| Description |
Gene expression data from mouse primary hepatocytes treated with Cav1 siRNA and TGFBeta
|
| Data processing |
The data were analyzed with a commercial software called JMP Genomics, version 6, from SAS. Gene expression profiling was performed using arrays of mouse Mogene-1_0-type from Affymetrix. A Custom CDF Version 16 with Entrez based gene definitions was used to annotate the arrays. The Raw fluorescence intensity values were normalized applying quantile normalization, RMA background correction and Medianpolish Probeset Summary
|
| |
|
| Submission date |
Sep 12, 2019 |
| Last update date |
Apr 10, 2020 |
| Contact name |
Carsten Sticht |
| Organization name |
University Heidelberg
|
| Department |
ZMF
|
| Street address |
Theodor-Kutzer-Ufer
|
| City |
Mannheim |
| ZIP/Postal code |
68169 |
| Country |
Germany |
| |
|
| Platform ID |
GPL19485 |
| Series (1) |
| GSE137339 |
Gene expression profiling of primary hepatocytes stimulated with TGF-β in the presence/absence of Caveolin-1 |
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