|
| Status |
Public on Sep 11, 2014 |
| Title |
Purified lung lavage macrophages WT, PND 3, biological replicate 3 |
| Sample type |
RNA |
| |
|
| Source name |
Wildtype, macrophages, specfic pathogen free facility
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57Bl/6N Tac
animal facility type: specfic pathogen free
genotype: Wildtype
age (postnatal day): 3
|
| Treatment protocol |
Mice are either wildtype or Scnn1b-transgenic and were harvested at post natal days 0, 3, 10, and 42. At PND 42 macrophages were also isolated from wildtype and Scnn1b-transgenic mice grown in a germ-free environment.
|
| Growth protocol |
Mouse lung lavage macrphages purified by negative selection through from a Lys-6G column; mice grown either under specific-pathogen-free conditions in micro-isolator cages or in a germ-free environment
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Magnetic activated cell sorting uas used to purify macrophages from lung lavage samples using Ant-Ly-6G MicroBeads (Miltenyi Biotec, MA). Ly-6G negative cells (macrophages) were pelleted, snap-frozen and stored at -80 C. Frozen macrophage pellets were lysed and homogenized in lysis buffer and QIAshedder (Qiagen). Genomic DNA was eliminated using gDNA eliminator spin columns; RNA was isolated using Purelink RNA mini kits (Invitrogen).
|
| Label |
biotin
|
| Label protocol |
12 ng of RNA was used to generate cDNA using Ovation Pico WTA system V2 kig (NuGEN Technologies, CA).
|
| |
|
| Hybridization protocol |
Hybridization was performed at 48 degrees using the GeneTitan plate system.
|
| Scan protocol |
Affymetrix GeneChip Scanner 3000 7G
|
| Description |
purified macrophages
|
| Data processing |
CEL files were imported into Partek Genomics Suite v6.5, and exon-level log2 intensities were obtained through RMA background correction, GC content and sequence adjustment, quantile normalization, and median polish probeset summarization. Gene-level log2 intensity were summarized as the mean of exon probesets.
MoGene-1_1-st-v1.r4.pgf
MoGene-1_1-st-v1.r4.mps
|
| |
|
| Submission date |
May 31, 2013 |
| Last update date |
Sep 12, 2014 |
| Contact name |
Hong Dang |
| E-mail(s) |
dangh@email.unc.edu
|
| Organization name |
UNC Chapel Hill
|
| Department |
CF Center
|
| Street address |
7027 Thurston-Bowles Bldg. CB#7248
|
| City |
Chapel Hill |
| State/province |
NC |
| ZIP/Postal code |
27599 |
| Country |
USA |
| |
|
| Platform ID |
GPL11533 |
| Series (2) |
| GSE47548 |
Gene expression in whole lung and pulmonary macrophages reflects the dynamic pathology associated with airway surface dehydration |
| GSE47551 |
mRNA expression data from either wild-type mice or Scnn1b-transgenic mice at post-natal days 0, 3, 10, and 42 |
|
