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Status |
Public on Oct 08, 2016 |
Title |
KO2-NI-1 |
Sample type |
RNA |
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Source name |
enSLiT07573 Knockout HeLa cells
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Organism |
Homo sapiens |
Characteristics |
infection: non-infected
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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.
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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.
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|
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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.
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Scan protocol |
Washed slides were scanned by Agilent's SureScan microarray scanner.
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Data processing |
The scanned images were anlyzed with Feature Extraction and text files were imported to GeneSpring GX to analyze differentially expressed transcripts.
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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 |
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