|
| Status |
Public on May 23, 2014 |
| Title |
Insulin-2 |
| Sample type |
RNA |
| |
|
| Source name |
hepatoma
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: hepatoma carcinoma cell
cell line: HepG2
tissue: liver
|
| Treatment protocol |
R-HepG2 cells were treated with 1 ml serum-free DMEM overnight, after which the cell culture medium was replaced with 1mL DMEM, 1 ml insulin or BI solution (total insulin concentration was identical, 40 ug/ml). The cells were incubated at 37 °C and 5% CO2 for 4 h and washed using phosphate-buffered saline (PBS, pH = 7.4) and 0.03% ethylene diamine tetraacetic acid (EDTA) to remove any residual complexes.
|
| Growth protocol |
R-HepG2 cells were cultured on tissue culture dishes in DMEM supplemented with 10% FCS, 50 mg/mL penicillin and 50 mg/mL streptomycin at 37 °C with 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using TRIZol Reagent (Life technologies) and purified using RNeasy Mini kit (QIAGEN), following manufacturers’ protocols.
|
| Label |
biotin
|
| Label protocol |
Total RNA were amplified, labeled, and purified using GeneChip® 3’IVT Express Kit (Affymetrix) according to the manufacturer’s instructions to obtain biotin labeled cRNA.
|
| |
|
| Hybridization protocol |
Labeled cDNA was hybridized to Affymetrix：GeneChip® PrimeView™ Human Gene Expression Array. Array hybridization and wash were performed using GeneChip Hybridization, Wash and Stain Kit (Affymetrix) in Hybridization Oven 645 (Affymetrix) and Fluidics Station (Affymetrix) by following the manufacturer’s instructions.
|
| Scan protocol |
Slides were scanned by GeneChip® Scanner 3000.
|
| Description |
HepG2 cells
|
| Data processing |
Slides were scanned by GeneChip® Scanner 3000 and Command Console Software 3.1 with default settings. Spike-in control transcripts were monitored to verify hybridization integrity.
|
| |
|
| Submission date |
May 22, 2014 |
| Last update date |
May 23, 2014 |
| Contact name |
Yun Xiao |
| E-mail(s) |
xy91269264@163.com
|
| Organization name |
Zhejiang University
|
| Street address |
Yuhangtang Road
|
| City |
Hangzhou |
| ZIP/Postal code |
310058 |
| Country |
China |
| |
|
| Platform ID |
GPL15207 |
| Series (2) |
| GSE57893 |
Towards epigenetic understanding and therapy of insulin resistance by intranuclear insulin [human cell line gene expression] |
| GSE57928 |
Towards epigenetic understanding and therapy of insulin resistance by intranuclear insulin |
|
