|
| Status |
Public on Apr 01, 2013 |
| Title |
Negative control shRNA transduced cells, not treated with isoproterenol, replicate 1 |
| Sample type |
RNA |
| |
|
| Source name |
Negative control shRNA transduced cells, not treated with isoproterenol
|
| Organism |
Mus musculus |
| Characteristics |
cell type: 3T3-L1 adipocytes
vector: Control shRNA
treatment: none
|
| Treatment protocol |
Mature adipocytes 9 days after induction of differentiation were transduced with adenoviruses carrying shRNA against TBLR1 or a control sequence at a multiplicity of infection (MOI) of 500. After 3 days, cells were preincubated in Krebs-Ringer buffer for 2 hrs, then cells were stimulated with 10 µM isoproterenol for 3 hrs.
|
| Growth protocol |
3T3-L1 preadipocytes were cultured in low (1g/l) glucose DMEM medium containing 10% fetal calf serum (FCS) and 1% penicillin/streptomycin (P/S) and differentiated into mature white adipocytes two days post confluency by the addition of 1 µg/ml insulin, 0.5 mM 3-isobutyl-1-methylxanthine (IBMX), 0.25 µM dexamethasone and 1/1000 volume ABP (50 mg/ml L-ascorbate, 1 mM biotin, 17 mM pantothenate) in high (4.5 g/l) glucose DMEM containing 10% FCS and 1% P/S
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNeasy Mini Kit (Quiagen)
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol.
|
| |
|
| Hybridization protocol |
Hybridization (16h x 45°C) was processed according to the standard Affymetrix protocol.
|
| Scan protocol |
Affymetrix GeneArray Scanner3000
|
| Description |
Gene expression data from 3T3-L1 adipocytes treated with negative control shRNA
|
| Data processing |
The data were analyzed with a commercial software called JMP Genomics, version 4, from SAS. Gene expression profiling was performed using arrays of Mouse430_2 -type from Affymetrix. A Custom CDF Version 13 with Entrez based gene definitions was used to annotate the arrays. The Raw fluorescence intensity values were normalized applying quantile normalization.
|
| |
|
| Submission date |
Jan 22, 2013 |
| Last update date |
Apr 01, 2013 |
| Contact name |
Carsten Sticht |
| Organization name |
University Heidelberg
|
| Department |
ZMF
|
| Street address |
Theodor-Kutzer-Ufer
|
| City |
Mannheim |
| ZIP/Postal code |
68169 |
| Country |
Germany |
| |
|
| Platform ID |
GPL14661 |
| Series (1) |
| GSE43658 |
Transcriptional co-factor TBLR1 controls lipid mobilization in white adipose tissue |
|
