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| Status |
Public on Dec 15, 2018 |
| Title |
3T3-L1 WT_d(+4)_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
3T3-L1 wildtype preadipocytes induced with MDI cocktail on day (+4)
|
| Organism |
Mus musculus |
| Characteristics |
cell type: 3T3-L1 adipocytes
time point: day (+4)
shRNA: wildtype
|
| Treatment protocol |
After cells reached confluence (day 0), cells were induced to differentiate by changing the culture medium to DMEM supplemented with MDI (0.5 mM 3-isobutyl-1-methyxanthine (Sigma),1 μM dexamethasone (Sigma) and 167 nM insulin (Sigma)) every two days.
|
| Growth protocol |
3T3-L1 Pre-adipocytes were cultured in DMEM supplemented with 10% newborn calf serum (Gibco), passaged at 70–80% confluence.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was prepared using TRIzol® Reagent (Invitrogen life technologies) following the manufacturer's recommendations. RNA clean-up was performed with RNasey Mini Kit(Qiagen p/n 74104). Quantification and quality of RNA was monitored with NanoDrop ND-1000
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 1 ug RNA using the Quick Amp Labeling Kit, One-Color (Agilent p/n 5190-0442) according to the manufacturer's instructions, followed byRNeasy Mini Kit (Qiagen p/n 74104). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
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| |
|
| Hybridization protocol |
1.65 ug of Cy3-labelled cRNA was fragmented at 60°C for 30 minutes in a reaction volume of 55ul containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 55ul of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Microarrays (G2534A) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed at room temperature with GE Wash Buffer 1 (Agilent) and GE Wash buffer 2 (Agilent) with 37°C GE, then dried immediately.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505C) using one color scan setting for 4x44k array slides.
|
| Description |
Gene expression in 3T3-L1 WT cells differentiated on day (+4)
|
| Data processing |
The scanned images were analyzed with Agilent Feature Extraction Software (v11.0.1.1).
|
| |
|
| Submission date |
Nov 01, 2018 |
| Last update date |
Dec 15, 2018 |
| Contact name |
fan yi |
| E-mail(s) |
[email protected]
|
| Organization name |
State Key Laboratory of Natural and Biomimetic Drugs
|
| Department |
Peking University
|
| Street address |
Xue Yuan Road 38
|
| City |
Beijing |
| ZIP/Postal code |
100191 |
| Country |
China |
| |
|
| Platform ID |
GPL11202 |
| Series (1) |
| GSE122054 |
Expression data from mouse preadipocyte 3T3-L1 and slincRAD-shRNA-8 cells |
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