|
Status |
Public on Apr 01, 2011 |
Title |
CD11b+ F4/80+ FOXP3+ rep3 |
Sample type |
RNA |
|
|
Source name |
spleen of C57 FOXP3 GFP mice
|
Organism |
Mus musculus |
Characteristics |
cell type: CD11b+ F4/80+ FOXP3+ strain: C57 FOXP3/GFP
|
Treatment protocol |
none
|
Growth protocol |
Freshly isolated Macrophages and MacRegs were derived from 4-6 month old C57 FOXP3/GFP mice through two consecutive rounds of fluorescent cell sorting using CD11b and F4/80 and GFP FOXP3. Freshly isolated CD4+Foxp3- and CD4+Foxp3+ cells were obtained from C57 FOXP3/GFP mice of the same age, through two consecutive rounds of fluorescent cell sorting. Three independent isolations were executed on separate days to obtain biological replicates.
|
Extracted molecule |
total RNA |
Extraction protocol |
RNA was extracted according to manufacturers instructionsusing the Rneasy MiniKit (Qiagen) , RNA was measured with the NanoDrop 1000 (Thermo Scientific) and integrity verified through Agilent Bioanalyzer 2100 using the nano chip.
|
Label |
cy3
|
Label protocol |
Agilent Low RNA Input Linear Amplification Kit PLUS was used incorporating cy3, according to manufacturers instructions
|
|
|
Hybridization protocol |
Agilent In Situ Hybridization Kit Plus was used according to manufacturers instructions, and labeled samples were hybridized to the Mouse whole genome 4X44k chip.
|
Scan protocol |
GE2-v5_95_Feb07 (Read Only), using Agilent Technologies Scanner G2505B US45102967
|
Description |
MacReg
|
Data processing |
Images were quantified using Agilent Feature Extraction Software (version A.9.5.1.1). Using Genespring GX11, the data was normalized to the 75th percentile and a baseline transformation to median of all samples was performed. Probes were filtered by percentile, removing those that fall beneath the 20th %. Probes were then filtered by flags removing those that did not have passing or marginal in 3 of the 6 samples. The two treatment populations (Mac Foxp3 neg and Mac Foxp3 pos were averaged and a t-test was performed to select genes that were significantly differentialy expressed between both populations. This list was further screened for a minimum of 2 fold change between the populations.
|
|
|
Submission date |
Feb 22, 2011 |
Last update date |
Apr 01, 2011 |
Contact name |
Cheryl E Myers |
E-mail(s) |
myers.cheryl@mayo.edu
|
Phone |
480-301-4056
|
Organization name |
Mayo Clinic College of Medicine
|
Department |
Immunology
|
Lab |
Cohen
|
Street address |
13400 E Shea Blvd
|
City |
Scottsdale |
State/province |
AZ |
ZIP/Postal code |
85259 |
Country |
USA |
|
|
Platform ID |
GPL7202 |
Series (1) |
GSE23793 |
Gene expression profile of Foxp3 expressing naturally ocurring regulatory macrophages from mice |
|