characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2014-06-20T17:01:59Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T18:20:20Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2604492
characteristics.characteristic: External Id
characteristics.text: University of Oxford
characteristics.characteristic: INSDC center name
characteristics.text: 2014-06-20T17:01:59Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T18:20:20Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS482447
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-2661:Exp2-Irf5-KO-000m-R1
characteristics.characteristic: Submitter Id
characteristics.text: 0 hour
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UO_0000032
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: M1 Macrophages
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CL_0000235
characteristics.characteristic: cell line
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: For the generation of M1 macrophages differentiated with GM-CSF, bone marrow of wild type and IRF5 knock out C57Bl6 mice was cultured in RPMI-1640 medium (PAA Laboratories) supplemented with recombinant mouse GM-CSF (20 ng/ml; Preprotech). Input samples were processed exactly as the other samples but without the immunoprecipication step. After 8 d, cells were washed with PBS and replated, then stimulated with lipopolysaccharide (LPS - 100 ng/ml; Alexis Biochemicals) for 0 or 2 h. 300 million GM-CSF derived M1 macrophages were used for the IRF5 ChIP experiment. Cells were fixed for 10 minutes with 1 % formaldehyde, quenched with 125 mM of Tris pH 7.5 and washed with ice-cold PBS. Nuclear lysates were isolated as described previously (De Santa et al. 2007) and sonicated with a Bioruptor (Diagenode) to obtain chromatin fragment sizes that average 500bp. Each lysate, but for the input one, was immunoprecipitated with 10 microgram of the following antibodies: IRF5 (Abcam; ab21689). ChIP was performed for each antibody as described previously (Ghisletti et al. 2010). ChIPped DNA was quantified with the Quant-iT dsDNA High Sensitivity Assay Kit (Invitrogen #Q33120). DNA yields ranged from 10-20 ng. 300 million GM-CSF derived M1 macrophages were used for the IRF5 ChIP experiment. Cells were fixed for 10 minutes with 1 % formaldehyde, quenched with 125 mM of Tris pH 7.5 and washed with ice-cold PBS. Nuclear lysates were isolated as described previously (De Santa et al. 2007) and sonicated with a Bioruptor (Diagenode) to obtain chromatin fragment sizes that average 500bp. Each lysate, but for the input one, was immunoprecipitated with 10 microgram of the following antibodies: IRF5 (Abcam; ab21689). ChIP was performed for each antibody as described previously (Ghisletti et al. 2010). ChIPped DNA was quantified with the Quant-iT dsDNA High Sensitivity Assay Kit (Invitrogen #Q33120). DNA yields ranged from 10-20 ng.
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: gene knock out
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0000506
characteristics.characteristic: genetic modification
characteristics.text: IRF5 knock out
characteristics.characteristic: genotype
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_10090
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: fresh specimen
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/OBI_0000971
characteristics.characteristic: specimen with known storage state
characteristics.text: C57Bl6
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0004472
characteristics.characteristic: strain
characteristics.text: Exp2-Irf5-KO-000m-R1
characteristics.characteristic: title