|
| Status |
Public on Oct 08, 2016 |
| Title |
KO2-Inf-1 |
| Sample type |
RNA |
| |
|
| Source name |
enSLiT07573 Knockout HeLa cells
|
| Organism |
Homo sapiens |
| Characteristics |
infection: Salmonella
|
| Treatment protocol |
The S. enterica serovar Typhimurium virulent strain used in this study was χ3306. A total of 1 × 105 HeLa cells in each well of a 12-well plate were challenged with Salmonella at a multiplicity of infection (m.o.i.) of 75, unless otherwise stated. The plate was centrifuged for 5 min at 500 ×g to enhance and synchronise infection. The cells were incubated for 1 h at 37°C to permit phagocytosis, and the free bacteria were removed by three washes with prewarmed phosphate-buffered saline (PBS). DMEM containing 10% heat-inactivated FBS and 100 μg/ml gentamicin was added and the cells were incubated for an additional 2 h at 37°C in a humidified incubator with 5% CO2. The cells were subsequently incubated with DMEM containing 10% heat-inactivated FBS and 10 μg/ml gentamicin at 37°C in a humidified incubator with 5% CO2 for the indicated period.
|
| Growth protocol |
HeLa TO cells were grown in Dulbecco’s modified Eagle’s medium (DMEM), supplemented with 10% foetal bovine serum (FBS) and antibiotics at 37°C in a humidified incubator with 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
To obtain RNA, RNAiso plus was added and followed by RNA isolation in accordance with the manufacturer’sinstructions, using chloroform and 2-propanol.
|
| Label |
Cy3
|
| Label protocol |
Total RNA(100 ng) was reverse transcribed and labeled by Low Input Quick Amp WT Labeling kit (Agilent technologies) according to the manufacturer's instructions.
|
| |
|
| Hybridization protocol |
Each 600 ng of labeled cRNA was hybridized to SurePrint G3 Human Gene Expression microarray 8x60K v3 at 65°C for 17 hours.
|
| Scan protocol |
Washed slides were scanned by Agilent's SureScan microarray scanner.
|
| Data processing |
The scanned images were anlyzed with Feature Extraction and text files were imported to GeneSpring GX to analyze differentially expressed transcripts.
|
| |
|
| Submission date |
Oct 07, 2016 |
| Last update date |
Oct 08, 2016 |
| Contact name |
Yayoi Fukuoka |
| E-mail(s) |
yayoi_fukuoka@agilent.com
|
| Organization name |
Agilent
|
| Street address |
Hachioji Takakuracho
|
| City |
Tokyo |
| ZIP/Postal code |
192-8510 |
| Country |
Japan |
| |
|
| Platform ID |
GPL21185 |
| Series (1) |
| GSE87754 |
Gene expression analysis of enSLiT KO cells against Salmonella infection |
|
