|
| Status |
Public on Mar 13, 2014 |
| Title |
Primary murine microglia - LPS Stimulated rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Wild-type CD-1 mice
|
| Organism |
Mus musculus |
| Characteristics |
age: P0-P1 pups
strain: CD1
cell type: primary microglia
stimulation: LPS
tissue: brain
|
| Treatment protocol |
Media was changed the night before to serum free media (DMEM/Pen-Strep only) and then stimulated with 100 ng/ml of LPS, 20 ng/ml of IL-4 or equivalent volume of PBS for 4 hours
|
| Growth protocol |
Primary Microglia from P0 CD-1 mice were cultured for 5-7 days in DMEM/FBS/Penn-Strep with 10 ng/ml of both M-CSF and GM-CSF
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated by using QIAzol and miRNeasy Minikit per supplier's protocol
|
| Label |
biotin
|
| Label protocol |
Biotin labeling was performed using the Ambion WT Expression Kit (Life Technologies, Grand Island, NY) according to the manufacturer's protocol, followed by the GeneChip WT Terminal Labeling and Controls Kit (Affymetrix, Santa Clara, CA).
|
| |
|
| Hybridization protocol |
The labeled, fragmented DNA was hybridized to the Mouse Gene 1.0 ST Array (Affymetrix, Santa Clara, CA) for 18 hours in a GeneChip Hybridization oven 640 at 45oC with rotation (60 rpm). The hybridized samples were washed and stained using an Affymetrix fluidics station 450.
|
| Scan protocol |
After staining, microarrays were immediately scanned using an Affymetrix GeneArray Scanner 3000 7G Plus (Affymetrix, Santa Clara, CA).
|
| Description |
Brains from pooled litter
|
| Data processing |
Raw Affymetrix CEL files were normalized to produce Entrez Gene-identifier-specific expression values using the implementation of the Robust Multiarray Average (RMA) in the affy package in the Bioconductor software suite and the Brainarray mogene10stv1entrezgcdf R package (version 13.0.0) (http://brainarray.mbni.med.umich.edu/Brainarray/Database/CustomCDF/13.0.0/entrezg.download/MoGene10stv1_Mm_ENTREZG_13.0.0.zip).
|
| |
|
| Submission date |
Jul 29, 2013 |
| Last update date |
Mar 13, 2014 |
| Contact name |
Tsuneya Ikezu |
| E-mail(s) |
tikezu@bu.edu
|
| Organization name |
Boston University School of Medicine
|
| Department |
BUSM - Department of Pharmacology and Experimental Therapeutics
|
| Lab |
Laboratory of Molecular NeuroTherapeutics
|
| Street address |
72 East Concord St
|
| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02118 |
| Country |
USA |
| |
|
| Platform ID |
GPL11078 |
| Series (2) |
| GSE49329 |
Integrated expression profiles of mRNA and miRNA in polarized primary murine microglia (mRNA) |
| GSE49331 |
Integrated expression profiles of mRNA and miRNA in polarized primary murine microglia |
|
