|
| Status |
Public on Nov 01, 2017 |
| Title |
T2 fraction of primary parenchymal lung fibroblasts of control 1 stimulated with TGF-β1 |
| Sample type |
RNA |
| |
|
| Source name |
Primary parenchymal lung fibroblasts, T2, Control 1, TGF-β1
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: Primary parenchymal lung fibroblasts
tissue: lung parenchyma
|
| Treatment protocol |
TGF-β1 stimulation: 24 hrs before the stimulation the cells were put in complete Ham's F12 medium containing 0.5% FCS. Afterwards, the lung fibroblasts were stimulated with 7.5 ng/ml TGF-β1 in complete Ham's F12 medium containing 0.5% FCS for 24 hrs. After the stimulation the cells were harvested for the Ago2-IP experiments.
|
| Growth protocol |
Primary lung fibroblasts were cultured in complete Ham's F12 medium containing 10% FCS. At passage 5, 17-20 million cells were collected for the Ago2-IP experiment.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The RNA in the Total (T) fraction and the immunoprecipitated (IP) fraction were isolated using miRNeasy Mini Kit and miRNeasy Micro Kit (both of Qiagen), respectively, according to manufacturer's protocol.
|
| Label |
Cy3
|
| Label protocol |
RNA of the T and IP fraction (50-80 ng) was labeled with cyanine 3 (Cy3) and cyanine 5 (Cy5) using the Two-Color Low Input Quick Amp Labeling Kit (Agilent Technologies) according to manufacturer's protocol Version 6.9.1.
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| |
|
| Hybridization protocol |
Dye-swap hybridizations were performed overnight using Gene Expression Hybridization Kit (Agilent Technologies)
|
| Scan protocol |
GE2_1200_Jun14 protocol was used for scanning the slide in the Agilent SureScan Microarray Scanner
|
| Description |
US45102948_257236310965_S01_GE2_1200_Jun14_1_4.txt
Primary parenchymal lung fibroblasts of control 1, stimulated with TGF-β1, harvested at passage 5, cRNA of T fraction after Ago2-IP was used for mRNA expression profiling
|
| Data processing |
Feature Extraction software version 12.0.1.1 was used to extract information from probe features from our microarray scan data.
|
| |
|
| Submission date |
Mar 14, 2017 |
| Last update date |
Nov 01, 2017 |
| Contact name |
Jennie Ong |
| E-mail(s) |
[email protected]
|
| Organization name |
University Medical Center Groningen
|
| Street address |
Hanzeplein 1
|
| City |
Groningen |
| ZIP/Postal code |
9713 GZ |
| Country |
Netherlands |
| |
|
| Platform ID |
GPL21185 |
| Series (2) |
| GSE86183 |
mRNA expression profiling after Ago2-immunoprecipitation (IP) in unstimulated and TGF-β1-stimulated primary parenchymal lung fibroblasts of two control subjects |
| GSE86186 |
miRNA expression data before and after TGF-β treatment and Ago2-associated transcripts in lung fibroblasts |
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