characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2017-10-31T17:01:52Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-08-22T09:54:55Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA104222111
characteristics.characteristic: External Id
characteristics.text: Department of Basic Sciences and Aquatic Medicine, Faculty of Veterinary Medicine, Norwegian University of Life Sciences
characteristics.characteristic: INSDC center name
characteristics.text: 2017-10-31T17:01:52Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-08-22T09:54:55Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1881129
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-5937:Sample 17
characteristics.characteristic: Submitter Id
characteristics.text: 7.2
characteristics.unit: year
characteristics.characteristic: age
characteristics.text: Norwegian Red
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/LBO_0000198
characteristics.characteristic: breed
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: macrophage
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CL_0000235
characteristics.characteristic: cell type
characteristics.text: cattle
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/LBO_0001036
characteristics.characteristic: common name
characteristics.text: Protocols: Six healthy Norwegian Red cows at the age of 4-7 years were used, blood samples of 500 ml were collected from the neck of each animal in sterile glass bottles with sodium citrate as an anticoagulant. Peripheral blood mononuclear cells (PBMC) were extracted by density gradient centrifugation (2210 × g, 30 min) on lymphoprep (Axis-Shield, Norway). CD14+ cells were extracted by positive selection of monocyte differentiation antigen CD14 using anti-human CD14 MACS MicroBeads (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany), according to the manufacturer’s instructions. CD14+ cells were grown in 6-well dishes at a density of 1.5 × 106 cells per well in RPMI medium supplemented with 10% FCS (Invitrogen, Carlsbad, USA). Cells were left over night at 37°C in an atmosphere with 5% CO2. The phenotypic morphology of cells, i.e. differentiation of monocytes into an early-stage adherent macrophage phenotype, was visualized and confirmed by phase contrast microscopy. For each individual animal the wells with macrophages were grouped into four classes with as equal number of wells and cells per class as possible. Two classes were infected with S. agalactiae strains ST103 and ST12, respectively, in a multiplicity of infection (MOI) of 1 (1 bacterium per cell, on average). The third cell class (positive control) was exposed to 1 mg/mL of lipopolysaccharides (LPS, rough strains) from Salmonella minnesota Re 595 (Re mutant, Sigma-Aldrich) and the last class was left uninfected (negative control). After 1 h of exposure, 1 % of penicillin/streptomycin (60 pg/mL penicillin and 100 µg/mL streptomycin) were added to prevent growth of remaining extracellular bacteria. The controls and the infected cells were treated equally. Inhibition of bacterial growth by antibiotics was verified by microscopy. Incubation was continued for one additional hour for LPS-exposed cells and for 5 more hours for bacteria infected and negative control cell classes, a total of 6 h incubation. Cells were centrifuged (400 × g, 5 min), pellet was washed with cold PBS buffer, snap frozen in liquid nitrogen and transferred to -70 oC for further storage. Mirvana Ambion kit NEBNext® Multiplex Small RNA Library Prep Set for Illumina®
characteristics.characteristic: description
characteristics.text: normal
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000461
characteristics.characteristic: disease
characteristics.text: wild type genotype
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0005168
characteristics.characteristic: genotype
characteristics.text: Bos taurus
characteristics.characteristic: organism
characteristics.text: Bos taurus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9913
characteristics.characteristic: organism
characteristics.text: blood
characteristics.characteristic: organism part
characteristics.text: Bos taurus
characteristics.characteristic: scientific_name
characteristics.text: female
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000383
characteristics.characteristic: sex
characteristics.text: Sample 17
characteristics.characteristic: title