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| Status |
Public on Jul 15, 2022 |
| Title |
HepG2 cell, ATF4 transfect sample for ATF4 overexpression condition |
| Sample type |
RNA |
| |
|
| Source name |
HepG2 cell, ATF4 overexpression condition
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HepG2
treatment: ATF4 overexpression
|
| Treatment protocol |
HepG2 cells were seeded on 6-well plates at 70–80% confluent density and cultured for 24 h in antibiotic-free DMEM. For leucine starvation, the cells were grown in minimum essential medium (Sigma-Aldrich) lacking leucine but supplemented with 10% (v/v) fetal bovine serum. Before starvation, the cells were rinsed twice with phosphate-buffered saline (Gibco, Waltham, MA, USA).For overexpression experiments, ATF4-overexpressing plasmid and pcDNA3.1 empty vector were transfected into HepG2 cells using Invitrogen Lipofectamine 2000 (Thermo Fisher Scientific) following the manufacturer's instructions. After 6 h, the culture medium was replaced with a complete medium and incubated for further 24 h.
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| Growth protocol |
Human hepatocellular carcinoma cells (HepG2) were obtained from the American Type Culture Collection (Rockville, MD, USA) and cultured in Dulbecco's modified Eagle's medium (DMEM; Sigma-Aldrich, St. Louis, MO, USA) containing 10% (v/v) fetal bovine serum (Sigma-Aldrich) and 1% (v/v) penicillin-streptomycin (Sigma-Aldrich) at 37 °C and 5% (v/v) CO2 atmosphere.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
For global miRNA expression profiling, total RNA (1000 ng) was labeled using the FlashtagTM RNA Labeling Kit.
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| |
|
| Hybridization protocol |
After washing and staining, array scanning was performed using GeneChip Hybridization and Affymetrix Gene Array Scanner 3000
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| Scan protocol |
After washing and staining, array scanning was performed using GeneChip Hybridization and Affymetrix Gene Array Scanner 3000
|
| Description |
Gene expression data of HepG2 cell under ATF4 overexpression condition
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| Data processing |
The data were analyzed with Transcriptome Analysis Console using Affymetrix default analysis settings and global scaling as normalization method.
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| |
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| Submission date |
Jul 14, 2022 |
| Last update date |
Jul 15, 2022 |
| Contact name |
Sihui Ma |
| E-mail(s) |
masihui@toki.waseda.jp
|
| Organization name |
The university of Tokyo
|
| Street address |
Yayoi 1-1-1
|
| City |
Tokyo |
| ZIP/Postal code |
1130032 |
| Country |
Japan |
| |
|
| Platform ID |
GPL16384 |
| Series (1) |
| GSE208228 |
Expression data from 1. leucine deprivation and 2. ATF4 overexpression on HepG2 cells |
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