|
| Status |
Public on Dec 31, 2018 |
| Title |
Control vs Doxycycline addition (NICD overexpression) |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Non-treated control of LS174T-tetON-NICD cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell line background: LS174T
genotype/variation: LS174T-tetON-NICD
treatment: un-treated control
|
| Treatment protocol |
un-treated control
|
| Growth protocol |
LS174T cells were maintained in minimum essential medium supplemented with 10% fetal calf serum, 1% penicillin-streptomycin, and 4 mM L-glutamine.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using RNA Bee Reagent (Tel Test Inc) following manufacturer's instructions
|
| Label |
Cy5
|
| Label protocol |
First and 2nd strand cDNA synthesis, and synthesis of aRNA was done by Amino Allyl aRNA kit (Ambion, using 1μg of total RNA as a starting material. 10μg of aRNA were subsequently labeled either by Cy5 Mono-reactive Dye (Amersham) or Cy3 Mono-reactive Dye (Amersham).
|
| |
|
| Channel 2 |
| Source name |
LS174T-tetON-NICD cells, Doxycycline for 24h
|
| Organism |
Homo sapiens |
| Characteristics |
cell line background: LS174T
genotype/variation: LS174T-tetON-NICD
treatment: Induced with Doxycycline (100ng/ml) for 24 h.
|
| Treatment protocol |
Human recombinant TNF-a was added at a concentration of 50ng/ml for 24h. Doxycycline was added at a concentration of 100ng/ml for 24 h.
|
| Growth protocol |
LS174T cells were maintained in minimum essential medium supplemented with 10% fetal calf serum, 1% penicillin-streptomycin, and 4 mM L-glutamine.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using RNA Bee Reagent (Tel Test Inc) following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
First and 2nd strand cDNA synthesis, and synthesis of aRNA was done by Amino Allyl aRNA kit (Ambion, using 1μg of total RNA as a starting material. 10μg of aRNA were subsequently labeled either by Cy5 Mono-reactive Dye (Amersham) or Cy3 Mono-reactive Dye (Amersham).
|
| |
|
| |
| Hybridization protocol |
Hybridization was done using 1μg of Cy5-labeled aRNA (Contol) and 1μg of Cy3-labeled aRNA (test) in the Hybridization Chamber (Takara Bio) for 16h, 37℃. After hybridization, slides were washed sequentially.
|
| Scan protocol |
3D-Gene Scanner (Toray Industries Inc., Tokyo, Japan) was used for scanning. Images were quantified using Extraction (Toray Industries Inc., Tokyo, Japan).
|
| Description |
QH37820
|
| Data processing |
F635 Median column as Red signal and F532 Median column as Green signal.Global normalized, background subtracted data were obtained from log2 of processed Green signal/processed Red signal.
|
| |
|
| Submission date |
Aug 02, 2018 |
| Last update date |
Dec 31, 2018 |
| Contact name |
Ryuichi Okamoto |
| E-mail(s) |
[email protected]
|
| Organization name |
Tokyo Medical and Dental University
|
| Street address |
Tokyo Medical and Dental University
|
| City |
Tokyo |
| State/province |
Please Select |
| ZIP/Postal code |
113-8519 |
| Country |
Japan |
| |
|
| Platform ID |
GPL13915 |
| Series (1) |
| GSE118048 |
Gene expression profile upon TNF-a stimulation, forced expression of NICD, or TNF-a stimulation under forced expression of NICD, in human colon carcinoma-derived LS174T cells |
|
