title: Illumina Genome Analyzer II sequencing; proj008_chipseq
design: proj008_chipseq
library_construction_protocol: For the generation of M1 macrophages differentiated with GM-CSF, bone marrow of wild type C57Bl6 mice was cultured in RPMI-1640 medium (PAA Laboratories) supplemented with recombinant mouse GM-CSF (20 ng/ml; Preprotech). After 8 d, cells were washed with PBS and replated, then stimulated with LPS (100 ng/ml; Alexis Biochemicals). 50, 100 and 300 million GM-CSF derived M1 macrophages were used for each Pol II, RelA and IRF5 ChIP experiment, respectively. 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 Each lysate was immunoprecipitated with 10 usc_20x_hg19_rCRSchrm.fa.bamg of the following antibodies: IRF5 (Abcam; ab21689), RelA (Santa Cruz; sc-372) and PolII (Santa Cruz; sc-899). ChIP was performed for each antibody as described previously (Ghisletti et al. 2010). ChIPped DNA w
attributes.tag: Experimental Factor: immunoprecipitate
attributes.value: RelA
attributes.tag: Experimental Factor: stimulus
attributes.value: lipopolysaccharide
attributes.tag: Experimental Factor: time post LPS stimulation
attributes.value: 2 hour