|
| Status |
Public on Jul 25, 2016 |
| Title |
MEF_Lats2WT_PDL90 |
| Sample type |
RNA |
| |
|
| Source name |
Embryonic fibroblast cells derived from LAT2+/+ C57BL/6J mice
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6J
tissue: embryonic fibroblasts
genotype: Lats2+/+
clone name: #112-K
pdl: 90
|
| Treatment protocol |
no treatment
|
| Growth protocol |
Mouse embyonic fibroblasts (MEFs) were cultured in DMEM + 10% inactivated fetal bovine serum (FBS), penicillin (100U/mL), and streptomycin (100μg/mL) at 37°C, 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from cultured cells with miRNeasy Mini Kit (Qiagen) and processed according to the manufacturer’s protocol. The quality of the RNAs was estimated by using the RNA 6000 Nano LabChip Kit (p/n 5065-4476) on the Agilent 2100 Bioanalyzer (G2940BA; Agilent Technologies, Santa Clara, CA).
|
| Label |
cy3
|
| Label protocol |
100 ng of aliquots of total RNA was used for making miRNA probes according to the Agilent protocol (ver. 1.6). Briefly, total RNA was dephosphorylated with calf intestine alkaline phosphatase (Takara Bio, Shiga, Japan), denatured with dimethyl sulfoxide, and labeled with pCp-Cy3 using T4 RNA ligase (Takara Bio, Shiga, Japan) using the miRNA Labeling Reagent and Hybridization Kit (5190-0408, Agilent).
|
| |
|
| Hybridization protocol |
Refer to Agilent miRNA hybridisation protocol. Probes were hybridized at 55°C for 20 hours with rotation. Then the slides were washed by Gene expression wash buffer 1 at room temperature for 5 minutes and by Gene expression wash buffer 2 at 37°C for 5 minutes.
|
| Scan protocol |
After hybridization and washing, the slides were scanned using an Agilent scanner (G2505C).
|
| Description |
Embryonic fibroblast cells derived from LAT2+/+ C57BL/6J mice
|
| Data processing |
Images were extracted using Agilent Feature Extraction software (v 10.5.1.1).
|
| |
|
| Submission date |
Nov 21, 2014 |
| Last update date |
Jul 25, 2016 |
| Contact name |
Daisuke Okuzaki |
| E-mail(s) |
[email protected]
|
| Phone |
+81-6-6879-4935
|
| Organization name |
Osaka univ.
|
| Department |
Immunology Frontier Research Center
|
| Lab |
Human Immunology （Single Cell Genomics)
|
| Street address |
Yamadaoka 3-1
|
| City |
Suita |
| State/province |
Osaka |
| ZIP/Postal code |
565-0871 |
| Country |
Japan |
| |
|
| Platform ID |
GPL10384 |
| Series (2) |
| GSE63532 |
miRNA array profiling of Lats2 deficient mouse embryonic fibroblasts |
| GSE63538 |
LATS2 regulates repressive epigenetic integrity via regulation of Polycomb repressive complex 2 |
|
