|
| Status |
Public on Feb 18, 2011 |
| Title |
GnRH treated LbetaT2 cells Expt2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
LbetaT2 gonadotrope cells
|
| Organism |
Mus musculus |
| Characteristics |
treatment: untreated
cell type: LbetaT2 gonadotrope cells
|
| Treatment protocol |
Cells were starved in DMEM containing 0.5% FBS, penicillin/streptomycin, and Gluta-max for 24 h then treated with 100 nM GnRH for 2 h
|
| Growth protocol |
Mouse LbetaT2 cells at passages 13-19 were cultured in monolayers with DMEM containing 10 % FBS, penicillin/streptomycin, and Gluta-max in a humidified 10 % CO2 atmosphere at 37˚C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was purified with RNA-Bee (Tel-test, TX) according to manufacturer’s protocol, the mRNAs were enriched from total RNA using the ribosomal RNA reduction kit (Invitrogen)
|
| Label |
Cy3
|
| Label protocol |
RNA from unstimulated cells was labeled with Cy3 and RNA from GnRH stimulated cells with Cy5 using the NCODE labelling kit following manufacturers instructions
|
| |
|
| Channel 2 |
| Source name |
LbetaT2 gonadotrope cells
|
| Organism |
Mus musculus |
| Characteristics |
cell type: LbetaT2 gonadotrope cells
treatment: 100 nM GnRH treated
|
| Treatment protocol |
Cells were starved in DMEM containing 0.5% FBS, penicillin/streptomycin, and Gluta-max for 24 h then treated with 100 nM GnRH for 2 h
|
| Growth protocol |
Mouse LbetaT2 cells at passages 13-19 were cultured in monolayers with DMEM containing 10 % FBS, penicillin/streptomycin, and Gluta-max in a humidified 10 % CO2 atmosphere at 37˚C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was purified with RNA-Bee (Tel-test, TX) according to manufacturer’s protocol, the mRNAs were enriched from total RNA using the ribosomal RNA reduction kit (Invitrogen)
|
| Label |
Cy5
|
| Label protocol |
RNA from unstimulated cells was labeled with Cy3 and RNA from GnRH stimulated cells with Cy5 using the NCODE labelling kit following manufacturers instructions
|
| |
|
| |
| Hybridization protocol |
Labeled probes were hybridized to NCODE miRNA microarrays in a Maui hybridization station for 24 h
|
| Scan protocol |
Arrays were scanned on a Molecular Devices Genepix 4000B
|
| Description |
miRNA expression
|
| Data processing |
mean of ratio (635/532) then miRNAs with signals < 2x background were excluded. Signals for the remaining miRNAs were median-centered and analyzed using Prism (Graphpad software) using a Z-test.
|
| |
|
| Submission date |
Feb 17, 2011 |
| Last update date |
Feb 18, 2011 |
| Contact name |
Nick Webster |
| E-mail(s) |
nwebster@ucsd.edu
|
| Phone |
858 534 6275
|
| Organization name |
UCSD
|
| Department |
Medicine
|
| Street address |
9500 Gilman Drive
|
| City |
La Jolla |
| State/province |
CA |
| ZIP/Postal code |
92093 |
| Country |
USA |
| |
|
| Platform ID |
GPL13189 |
| Series (1) |
| GSE27398 |
Effect of GnRH on miRNA expression in LbetaT2 cells |
|
