Rattus norvegicus
Protocols: At E14.5, a single cell suspension was obtained from pooled lungs from either control or nitrofen exposed rat fetuses by trypsinization (0.25% Trypsin-EDTA, ThermoFisher, Waltham, MA) for 20 minutes at 37°C humidified chamber. Cells were spun down by centrifugation for 5 minutes at 800g and the pellet was resuspended in DMEM supplemented with 10% FBS and subjected to three serial depletion of fibroblasts by incubation for 1 hour each following an established protocol [Caniggia 1991]. Groups were assigned randomly. Treatment with EVs is with 10% (by volume) Total RNA was extracted using Nucleospin miRNA kit (Macherey-Nagel, Düren, Germany), following supplier recommended protocols. We used an automated NEBNext Ultra II Directional with polyA isolation (New England BioLabs) using the Agilent NGS Workstation (Agilent Technologies) as per manufacturer's protocol. Briefly, 250 ng of total RNA spiked-in with ERCCs (External RNA Controls Consortium, Ambion) as per manufacturer's protocol was used to generate cDNA.
ENA-FIRST-PUBLIC
2021-03-01T12:16:09Z
ENA-LAST-UPDATE
2020-04-11T12:25:22Z
External Id
SAMEA6663519
INSDC center name
Developmental and Stem Cell Biology Program, Peter Gilgan Centre for Research and Learning, The Hospital for Sick Children
INSDC first public
2021-03-01T12:16:09Z
INSDC last update
2020-04-11T12:25:22Z
INSDC status
public
Submitter Id
E-MTAB-8921:Sample 22
age
14.5
broker name
ArrayExpress
cell type
epithelial cell of lung
common name
Norway rat
developmental stage
pseudoglandular stage
disease
normal
fraction
cell
genotype
wild type genotype
organism part
lung epithelium
sample name
E-MTAB-8921:Sample 22
scientific_name
Rattus norvegicus
sex
not available
strain
Sprague-Dawley