|
| Status |
Public on Feb 11, 2010 |
| Title |
C-peptide_18hrs_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
HK-2 cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HK-2
|
| Treatment protocol |
For microarray analyses, cells were cultured in 5mM-DMEM/F12 without supplements (control) or with 5nM C-peptide for either 18h or 48h. Alternatively, cells were treated with 2ng/ml TGF-b1 either alone or in combination with C-peptide for 48h. All treatments were performed in triplicate to yield 18 flasks, and RNA from each flask hybridized to a separate chip to give an n of 3 for each of 6 treatments. Flasks were subjected to identical media changes and cells cultured for identical periods in media without supplements. In all experiments, cells were serum-starved overnight before agonist addition. Treatments were initiated such that 18h and 48h incubation periods ended coincidentally and all RNA was prepared at this point.
|
| Growth protocol |
HK-2 cells (passages 18-30) were maintained in DMEM/Hams F12 (DMEM/F12) (17.5mM glucose), supplemented with 10% fetal calf serum (FCS), glutamine (2mM), penicillin (100 IU/ml) and streptomycin (100ng/ml). Cells were cultured at 37oC in a humidified atmosphere of 5% CO2 in air. Prior to treatment, cells were cultured in DMEM/F12 low glucose (5mM) (5mM-DMEM/F12) for 48h.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was prepared by acid-guanidinium extraction using a Genelute mammalian total RNA miniprep kit (Sigma-Aldrich, Poole, UK) following the manufacturer’s instructions.
|
| Label |
biotin
|
| Label protocol |
Standard Illumina hybridization protocol performed by Geneservice (Cambridge, UK)
|
| |
|
| Hybridization protocol |
Standard Illumina hybridization protocol performed by Geneservice (Cambridge, UK)
|
| Scan protocol |
Standard Illumina scanning protocol performed by Geneservice (Cambridge, UK)
|
| Description |
HK-2 cells cultured 18h with 5nM C-peptide
|
| Data processing |
The array intensity data were analysed using Illumina GenomeStudio Gene Expression Module (v1.1.1) (Illumina Cambridge, UK) for visualisation and normalisation. The quantile normalisation method was used for all analyses and average background correction performed within the Beadstudio software.
|
| |
|
| Submission date |
Feb 09, 2010 |
| Last update date |
Feb 12, 2010 |
| Contact name |
Nigel John Brunskill |
| E-mail(s) |
njb18@le.ac.uk
|
| Phone |
+44 116 2588043
|
| Organization name |
University of Leicester
|
| Department |
Infection Immunity and Inflammation
|
| Street address |
University Road
|
| City |
Leicester |
| State/province |
Leics |
| ZIP/Postal code |
LE1 9HN |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL6884 |
| Series (1) |
| GSE20247 |
C-peptide and/or transforming growth factor beta 1 effect on human proximal tubular cell line |
|
