|
| Status |
Public on Mar 07, 2018 |
| Title |
C-E-2 |
| Sample type |
RNA |
| |
|
| Source name |
human corneal epithelial cell line, control
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: HCEC
medium formulation: KSFM with bovine pituitary extract (BPE, 25 ug/mL), epidermal growth factor (EGF, 50 ng/ml), penicillin and streptomycin
2 day pre-treatment: EtOH
6hr treatment: BSA
|
| Treatment protocol |
After reaching confluence, cells were rinsed with PBS and cultured in medium containing DMEM/F12 with 10% FBS, 10 ng/ml EGF, penicillin and streptomycin and either dihydrotestosterone (DHT, 10 nM) or ethanol control for 2 days. After this time period, cells were incubated in serum-free DMEM/F12 and exposed to vehicle (1% bovine serum albumin [BSA]), or LPS (15 μg/ml) and LBP (150 ng/ml), for six hours.
|
| Growth protocol |
Cells were cultured in keratinocyte serum-free medium (KSFM) supplemented with bovine pituitary extract (BPE; 25 ug/mL HCEC and HCjE, 50 ug/mL HMGEC), 5 ng/mL epidermal growth factor (EGF), penicillin and streptomycin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted wth Rneasy Mini Kit(Qiagen) per manufacturer's recommendations. Quality control and quantification were performed with Agilent Bioanalyser. Quality and quantity were checked again before subjected to microarray by Asuragen.
|
| Label |
Biotin
|
| Label protocol |
Biotinylated cRNA were prepared using a MessageAmp II-based protocol (Ambion Inc., Austin, TX) and one round of amplification.
|
| |
|
| Hybridization protocol |
Standard Illumina hybridization protocol
|
| Scan protocol |
Standard Illumina scanning protocol
|
| Data processing |
Data were were cubic spline normalized using Illumina Bead Studio software. After normalization the data were further processed by changing all negative values to zero and then adding 16 to each intensity.
|
| |
|
| Submission date |
Mar 06, 2018 |
| Last update date |
Mar 07, 2018 |
| Contact name |
David A. Sullivan |
| E-mail(s) |
david.sullivan@schepens.harvard.edu
|
| Phone |
617-912-0287
|
| Organization name |
Schepens Eye Research Institute
|
| Lab |
David A. Sullivan
|
| Street address |
20 Staniford St.
|
| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02114 |
| Country |
USA |
| |
|
| Platform ID |
GPL10558 |
| Series (1) |
| GSE111496 |
The effects of testosterone and lipopolysaccharide on gene expression in immortalized ocular surface and adnexal cells |
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