|
Status |
Public on Oct 01, 2014 |
Title |
2.SIRT1 young |
Sample type |
RNA |
|
|
Source name |
SIRT1 young
|
Organism |
Mus musculus |
Characteristics |
age: young genotype: SIRT1F/F cell type: microglia genetic background: BL6 and 129/Sv
|
Treatment protocol |
Adult microglia were isolated from SIRT1F/F or LysM-Cre/SIRT1F/F mice as described with some modifications. Brains were placed in a culture dish with enzyme solution and the digested brains were triturated with a 20-ml pipette and passed through 70-μm filter. After centrifugation, cells were incubated with anti-myelin microbeads, centrifuged, resuspendend and washed. Myelin-negative fractions were collected and further processed by incubating with anti-CD11b microbeads at 4°C for 15 min. After washing, the resuspended myelin negative fraction was passed through an autoMACS Pro Separator, and then the CD11b-positive population was separated from CD11b-negative population. Cells were centrifuged, and the pellets were lysed with RNA lysis buffer (QIAGEN) for microarray or quantitative RT-PCR.
|
Extracted molecule |
total RNA |
Extraction protocol |
RNA was extracted with standard methods
|
Label |
Biotin
|
Label protocol |
Biotinylated cRNA were prepared with Illumina kit for Illumina arrays
|
|
|
Hybridization protocol |
Standard Illumina hybridization protocol
|
Scan protocol |
Standard Illumina scanning protocol
|
Data processing |
The data were log2 transformed and normalised using quantile normalisation in R
|
|
|
Submission date |
Apr 02, 2014 |
Last update date |
Oct 01, 2014 |
Contact name |
Giovanni Coppola |
E-mail(s) |
gcoppola@ucla.edu
|
Phone |
310-794-4172
|
Organization name |
UCLA
|
Department |
Psychiatry and Neurology
|
Lab |
Neurogenetics
|
Street address |
1524 Gonda, 695 Charles Young Drive South
|
City |
Los Angeles |
State/province |
CA |
ZIP/Postal code |
90095 |
Country |
USA |
|
|
Platform ID |
GPL6885 |
Series (1) |
GSE56452 |
SIRT1 deficiency in microglia contributes to cognitive decline in aging and neurodegeneration via epigenetic regulation of IL-1beta |
|