|
| Status |
Public on May 01, 2020 |
| Title |
eosinophils, mock infected, biological replicate 5 |
| Sample type |
RNA |
| |
|
| Source name |
bone marrow cultured in eosinophil growth media for 14 days
|
| Organism |
Mus musculus domesticus |
| Characteristics |
strain: C57BL/6
cell type: eosinophils
gender: Female
age: 6 weeks old
influenza strain: Mock
|
| Treatment protocol |
On day 14 of differentiation, BMdEos were pulsed for 1 h with MOI 0.1 PR8. Cells were washed in PBS and cultured for a further 23 h at a density of 1 million live cells/mL in media supplemented with 1 µg/mL TPCK-Trypsin and 5 ng/mL mouse GM-CSF. At six hours prior to harvest, 1 µg/ml GolgiPlug (BD Biosciences) was added to the cells to reduce the release of endogenous RNases.
|
| Growth protocol |
Eosinophils were derived from female C57BL/6 bone marrow by culturing for 14 days in eosinophil growth media.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted from mock or PR8-pulsed BMdEos using RNeasy Mini (Qiagen Inc., Germantown, MD, USA) with β-mercaptoethanol added to the RLT buffer, as directed by the manufacturer, and eluted into RNase-free water.
|
| Label |
biotin
|
| Label protocol |
Sense target cDNA was prepared from 100ng of total RNA using the Ambion® WT Expression Kit (Life Technologies P/N 4411973). cDNA was fragmented and bioton-labeled using the Affymetrix® WT Terminal Labeling Kit (Affymetrix P/N 900671).
|
| |
|
| Hybridization protocol |
Following fragmentation, 5.5ug of cDNA were hybridized for 16 hr at 45C on GeneChip Mouse Gene 2.0 ST arrays. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GCS 3000 7G.
|
| Description |
Gene expression data from mouse bone marrow derived eosinophils, mock infected
|
| Data processing |
Probe signals were quantile normalized and summarized by the PLIER algorithm using Affymetrix Expression Console software v1.1.
|
| |
|
| Submission date |
Nov 18, 2019 |
| Last update date |
May 02, 2020 |
| Contact name |
Robert Rooney |
| E-mail(s) |
rrooney1@uthsc.edu
|
| Phone |
9012875364
|
| Organization name |
University of Tennessee Health Science Center
|
| Department |
Pediatrics
|
| Lab |
Robert Rooney
|
| Street address |
50 N. Dunlap, Room 366R, LeBonheur CFRC
|
| City |
Memphis |
| State/province |
TN |
| ZIP/Postal code |
38103 |
| Country |
USA |
| |
|
| Platform ID |
GPL16570 |
| Series (1) |
| GSE140580 |
Gene Expression Data from Influenza A Virus-Infected and Mock-infected Mouse Eosinophils |
|
