title: RNAseq of a chicken lung epithelial cell line (cLEC-213) upon treatment with butyrate and/or infection with low pathogenic avian influenza
design: RNAseq of a chicken lung epithelial cell line (cLEC-213) upon treatment with butyrate and/or infection with low pathogenic avian influenza
library_name: Sample 8_p
library_construction_protocol: 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.
identifiers.namespace: European Bioinformatics Institute
identifiers.id: E-MTAB-14346:Sample 8_p