|
| Status |
Public on Aug 08, 2021 |
| Title |
pro-BMP9 treatment, PAEC line 2 |
| Sample type |
RNA |
| |
|
| Source name |
pulmonary artery endothelial cells
|
| Organism |
Homo sapiens |
| Characteristics |
Sex: male
age: 40
treatment: pro-BMP9, containing 0.4 ng/ml of final growth factor domain
|
| Treatment protocol |
All ligands (at a final concentration of 0.4 ng/ml growth factor concentration) were added to the serum-starved cells. The 6-well plates were returned into CO2 incubator at 37 ºC for 5 hours before cells were harvested for RNA extraction.
|
| Growth protocol |
PAECs were seeded in 6-well plates and grown in EGM-2 with 10% FBS to 90% confluency. Cells were then put into EGM-2 with 0.1% FBS for 16-18 hours before the treatments applied.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNeasy Mini Kit from Qiagen following manufacturer's instruction
|
| Label |
biotin
|
| Label protocol |
100 ng total RNA was amplified along with inline PolyA spike in control RNA, using the WT PLUS amplification kit (Affymetrix). Successfully amplified samples were labelled using the GeneChip WT terminal labelling kit (Affymetrix) Using the in line hybridization controls.
|
| |
|
| Hybridization protocol |
Plate arrays were processed on the GeneTitan instrument (Affymetrix) using the GeneTitan Hybridization, Wash and Stain kit (Affymetrix). Samples were hybridized to the array, washed, stained and scanned using the array specific parameters provided by Affymetrix.
|
| Scan protocol |
Automated protocol from the GeneTitan, according to the automated settings provided for the array
|
| Data processing |
The processing of the data was carried out in R, using the package Oligo and the normalisation method used is RMA (Robust Multichip Analysis). The comparisons were performed using the limma package
|
| |
|
| Submission date |
Aug 09, 2018 |
| Last update date |
Aug 08, 2021 |
| Contact name |
Wei Li |
| E-mail(s) |
wl225@cam.ac.uk
|
| Phone |
+44 1223 761304
|
| Organization name |
University of Cambridge
|
| Department |
Medicine
|
| Street address |
Hills Road
|
| City |
Cambridge |
| ZIP/Postal code |
CB2 0QQ |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL17692 |
| Series (1) |
| GSE118353 |
Pro-BMP9 signalling in human pulmonary arterial endothelial cells after 5 hours treatment |
|
