2024-08-29
ENA first public
ERC000011
ENA-CHECKLIST
INRAE
INSDC center name
public
INSDC status
ERS20862484
SRA accession
E-MTAB-14346:Sample 15
Submitter Id
ArrayExpress
broker name
CLEC-213
cell_line
epithelial cell
cell_type
not collected
collection date
chicken
common name
Protocols: The spontaneously immortalized chicken respiratory epithelial cell line CLEC213 has previously been obtained from the lungs of histocompatible B21/B21 white leghorn layer chickens (PA12). Cells were cultured in Dulbecco’s Modified Eagles’s Medium (DMEM) supplemented with 10% FCS (Gibco, UK), 2 mM L-Glutamine (Gibco, UK), and 100 UI/ml penicillin and 100 μg/ml streptomycin (PS, Gibco, UK) at 41°C in a 5% CO2 incubator.  AIV infection experiments using CLEC213 were performed to assess the effects of butyrate pre-treatment for 16h on viral replication and innate immune gene regulation. To this end, subconfluent CLEC213 monolayers (3x105 cells/well in 6-wells plates) were pre-treated with butyrate (3 mM) for 16h and then infected with the AIV strains A/Mallard/Marquenterre/Z237/83 (H1N1) or A/Turkey/Italy/977/1999 (H7N1) at a multiplicity of infection (MOI) of 0.1 or 0.01 by incubating the cells with a virus dilution prepared in serum-free DMEM for 1 h at 37 °C. Following this 1 h adsorption period, cells were washed with PBS and then overlayed with serum-free DMEM containing TPCK-treated trypsin (0.4 μg/ml final). Total RNA was extracted using the NucleoSpin RNA II kit (Macherey-Nagel, Germany) with rDNAse treatment to eliminate contaminant DNA. RNA quality was checked by the Bioanalyzer 2100 system using the RNA Nano 6000 Assay kit (Agilent Technologies, CA, USA).  The NEBNext® Ultra™ II RNA Library Prep Kit for Illumina® was used to prepare the library. RNA with PolyA selection.
description
adult
developmental stage
avian influenza
disease
B21/B21
genotype
not collected
geographic location (country and/or sea)
Mock
infect
not applicable
isolate
Gallus gallus
organism
Gallus gallus
organism
lung
organism part
Gallus gallus
scientific_name
Sample 15
title