|
| Status |
Public on Jul 19, 2024 |
| Title |
TGFB1_24h_2 |
| Sample type |
RNA |
| |
|
| Source name |
hepatocyte isolated from male DBA/2J mice (8-12 weeks old)
|
| Organism |
Mus musculus |
| Characteristics |
treatment: 5ng/mL recombinant TGF-β1
time: 24h
|
| Treatment protocol |
5ng/mL recombinant TGF-β1 was added to serum-free culture medium for 1, 6, 24 and/or 48 hours. Control conditions included cells maintained for the same period in a serum-free medium without TGF-beta1.
|
| Growth protocol |
Primary hepatocytes were isolated from livers of male DBA/2J mice (8-12 weeks old) using collagenase perfusion. Hepatocytes were plated on collagen-coated 6-well plates at a density of 3x105 cells/well in Williams' medium E supplemented with 10% FBS, 2mM L-glutamine, 1% penicillin/streptomycin and 100nM dexamethasone. Hepatocytes were allowed to attach, and medium was exchanged after 4h with Williams medium E supplemented with 2mM L-glutamine and 1% penicillin/streptomycin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
After cytokine treatment, cultured cells were washed twice with PBS. Next, total RNA was isolated using the Total RNA Kit, peqGOLD (Cat No. 13-6834-02) or the InviTrap®Spin Universal RNA Mini Kit from Stratec (1060100300, Birkenfeld, Germany) according to manufacturers´ instructions. RNA concentration was measured with the Infinite M200 microplate reader (Tecan, Switzerland).
|
| 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
|
| Data processing |
The data were analyzed with R (v 4.2.2.). Gene expression profiling was performed using arrays of Mouse430_2 from Affymetrix. A Custom CDF Version 25 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 |
Mar 08, 2024 |
| Last update date |
Jul 19, 2024 |
| Contact name |
Carolina Delatorre |
| E-mail(s) |
carolina.delatorre@medma.uni-heidelberg.de
|
| Organization name |
University Heidelberg
|
| Street address |
Theodor-Kutzer-Ufer 1-3
|
| City |
Mannheim |
| ZIP/Postal code |
68167 |
| Country |
Germany |
| |
|
| Platform ID |
GPL34289 |
| Series (1) |
| GSE261215 |
Transcription profiling by array of mouse hepatocytes following treatment with growth factor beta 1 in a time series |
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