|
| Status |
Public on Jul 11, 2023 |
| Title |
MDCK Mix culture_2 |
| Sample type |
RNA |
| |
|
| Source name |
MDCK-pTR GFP-RasV12 cells co-cultured with cells at a ratio of 1:100
|
| Organism |
Canis lupus familiaris |
| Characteristics |
culture condition: mixed culture
|
| Treatment protocol |
MDCK-pTR GFP-RasV12 cells or a 10:1 mix culture of MDCK and MDCK-pTR GFP-RasV12 cells were cultured at a density of 1.2×107 cells on collagen-coated 10 cm dishes (CORNING). After incubation with tetracycline for 24 h at 37 ºC in a humidified incubator with 5% CO2, GFP-positive RasV12 cells was separated with an analytical flow cytometer.
|
| Growth protocol |
MDCK and MDCK-pTR GFP-RasV12 cells were cultured in Dulbecco's Modified Eagle Medium, High Glucose supplemented with 10% Fetal Bovine Serum and penicillin/Streptomycin at 37 ºC in a humidified incubator with 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from MDCK-pTR GFP-RasV12 mCherry-β-catenin Δ131 cells using ISOGEN II (NIPPON GENE Co., Ltd).
|
| Label |
Cy3
|
| Label protocol |
For cRNA amplification and labeling, Agilent Low Input Quick Amp Labeling Kit (Agilent Technologies) were used following the manufacter’s instructions.
|
| |
|
| Hybridization protocol |
1.5 ug of Cy3-labelled cRNA was fragmented at 60°C for 30 minutes in a Agilent fragmentation buffer and was hybridized at 65°C for 17 hr using Gene Expression Hybridization Kit.
|
| Scan protocol |
Microarray slides were scanned using an Agilent SureScan (G4900DA).
|
| Description |
Gene expression after 24h of tetracycline treatment
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software (Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
|
| |
|
| Submission date |
Jul 06, 2023 |
| Last update date |
Jul 11, 2023 |
| Contact name |
Shunsuke Kon |
| E-mail(s) |
kon44@rs.tus.ac.jp
|
| Phone |
+81-4-7121-4053
|
| Organization name |
Tokyo University of Science
|
| Department |
Research Institute for Biomedical Sciences
|
| Lab |
Division of Cancer Biology
|
| Street address |
2669 Yamazaki
|
| City |
Noda-shi |
| State/province |
Chiba |
| ZIP/Postal code |
278-0022 |
| Country |
Japan |
| |
|
| Platform ID |
GPL15379 |
| Series (1) |
| GSE236658 |
Gene expression signatures of MDCK-pTR GFP-RasV12 cells cultured alone or mixed with MDCK cells |
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