|
| Status |
Public on Mar 30, 2015 |
| Title |
sirt1_KO 2nd_neurosphere _6DIV_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
2nd neurosphere from sirt1 KO adult mice,cultured for 6 days
|
| Organism |
Mus musculus |
| Characteristics |
strain: 129/ICR
genotype/variation: sirt1 KO
tissue: hippocampus
developmental stage: 7 to 9 weeks
gender: female
cell type: secondary neurospheres
|
| Treatment protocol |
WT and KO neurospheres were cultured for 6 days before harvest. DMSO- and resveratrol- treated neurospheres were cultured for 5 days followed by 24 hours DMSO and 10 mm resveratrol treatment.
|
| Growth protocol |
Adult hippocampal neural stem cells were isolated by gradient centrifugation, seeded low adhension 6-well plate with Neurobasal A supplemented with B27 minus retinyl acetate, glutamax, penicillin/streptomycin, bFGF (10ng/ml), EGF(10ng/ml), PDGFbb(10ng/ml) and heparin (2 mg/ml). 7 to 9 days after seeding, primary neurospheres are dissociated and seeded again into low adhension 6-well plate at 5*10^4 cells/ml with NeurobasalA supplemented with B27 minus retinyl acetate, glutamax, penicillin/streptomycin, bFGF (20ng/ml), EGF(20ng/ml), and heparin (2 mg/ml).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was prepared with Trizol reagent (Invitrogen) according to the manufacturer's protocol.
|
| Label |
Cy3
|
| Label protocol |
Total RNA was amplified and labeled by Low input Quick Amp Labeling Kit, One-Color (Cat#5190-2305, Agilent technologies), followed the manufacturer's instructions.
|
| |
|
| Hybridization protocol |
Each slide was hybridized with 1.65 mg Cy3-labeled cRNA using Gen Expression Hybridization Kit (Cat#5188-5242, Agilent technologies)in Hybridization Oven (Cat#G2545A, Agilent technologies) at 65°C, 10rpm for 17 hours. Then slides were washed in staining dishes (Cat#121, Thermo Shandon) with Gene expression Wash Buffer Kit (Cat#5188-5327, Agilent technologies).
|
| Scan protocol |
Slides were scanned by Agilent Microarray Scanner (Cat#G2565CA, Agilent technologies) with default settings. Dye channel: Green, scan resolution=5 mm, PMT 100%, 10%, 16 bit.
|
| Description |
Gene expression profile of 2nd neurospheres from sirt1 KO mice hippocampus
|
| Data processing |
The scanned images were analysed with Feature Extraction software 10.7 (Agilent technologies). Raw data were normalized by Quantile algorithm using Gene Spring Software 11.0 (Agilent technologies)
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| |
|
| Submission date |
Jul 21, 2012 |
| Last update date |
Mar 30, 2015 |
| Contact name |
Chenyan Ma |
| Organization name |
UC Berkeley
|
| Street address |
230E Li Ka Shing Center
|
| City |
Berkeley |
| State/province |
CA |
| ZIP/Postal code |
94720 |
| Country |
USA |
| |
|
| Platform ID |
GPL7202 |
| Series (1) |
| GSE39551 |
Gene expression profiles of adult neural stem cells with loss-of-function and gain-of-function of SIRT1 |
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