|
| Status |
Public on Dec 05, 2019 |
| Title |
Steady State, LMYC KO2, Biological Rep 4 |
| Sample type |
RNA |
| |
|
| Source name |
FACS purified splenic cDC1
|
| Organism |
Mus musculus |
| Characteristics |
tissue: spleen
cell type: cDC1
genotype: MyclGFP/GFP
|
| Treatment protocol |
Spleens were minced and digested with stirring for 30 min at 37ºC in 5ml of IMDM plus 10% FBS with 30U/ml DNaseI (Sigma-Aldrich) and 250ug/ml collagenase B (Roche). For removal of debris, cells were passed through a 70-µm filter before red blood cells were lysed with ammonium chloride–potassium bicarbonate lysis buffer. For experiments with poly IC activation mice were injected intraperitoneally with PBS or PBS containing 100 micrograms of poly IC. For experiments with MYCL defieicnt mice, poly IC treatment lasted 5 hours prior to euthanization. For experiments with MXD1 defieicnt mice, poly IC treatment lasted 18 hours prior to euthanization.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Column extraction of total RNA was performed acording to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cDNA were prepared according to the Affymetrix WT Pico reagent kit from 100pg-2ng total RNA.
|
| |
|
| Hybridization protocol |
Following fragmentation and labeling, 2.5 ug of cDNA were hybridized onto the Gene 1.0 ST arrays and placed in GeneChip Hybridization Oven 640 for 18 hr at 45C. Gene Chip’s were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip 7G 3000 Scanner.
|
| Description |
Gene expression data at steady state in vivo
|
| Data processing |
The data was analyzed with the Affymetrix GeneChip Command Console. Microarray expression data was processed using Command Console (Affymetrix, Inc) and the raw (.CEL) files generated were analyzed using Expression Console software with Affymetrix default RMA Gene analysis settings.(Affymetrix, Inc). Probe summarization (Robust Multichip Analysis, RMA), quality control analysis, and probe annotation were performed according to recommended guidelines (Expression Console Software, Affymetrix, Inc.).
|
| |
|
| Submission date |
Dec 04, 2019 |
| Last update date |
Dec 05, 2019 |
| Contact name |
David Alexander Anderson III |
| E-mail(s) |
[email protected]
|
| Organization name |
Washington University School of Medicine
|
| Department |
Pathology and Immunology
|
| Lab |
Murphy
|
| Street address |
660. S Euclid Ave., Campus Box 8118
|
| City |
St. Louis |
| State/province |
Missouri |
| ZIP/Postal code |
63110 |
| Country |
USA |
| |
|
| Platform ID |
GPL6246 |
| Series (1) |
| GSE141492 |
The MYCL and MXD1 transcription factors regulate the fitness of murine dendritic cells |
|
