Expression Data from BALB/c and Stat6-deficient bone marrow derived macrophages (BMDM). Mus musculus

We used microarrays to find Stat6 dependent genes in control and IL-4 exposed bone marrow derived macrophages. Alternatively activated macrophages (AAM) accumulate in tissues during Th2-associated immune responses like helminth infections and allergic disorders. These cells possess potent inhibitory activity against T cells. The differentiation of AAM depends on IL-4/IL-13-mediated activation of the transcription factor Stat6. Stat6 is also required in AAM to induce several genes, such as YM1, FIZZ1 and Arginase1. Overall design: Macrophages were differentiated from bone marrow cells in supplemented RPMI 1640 for 7 days in the presence of 10% supernatant from the M-CSF producing fibroblast cell line L929. After removal of nonadherent cells, macrophages were detached using 5mM EDTA treatment, washed, counted, and replated at a density of 10x6 cells/mL. Macrophages were cultured for 24 hours in medium (control) or in the presence of IL-4 (10mg/mL). F4/80 positive macrophages were sorted with a purity of 99% and RNA was isolated (Trizol/Chloroform) according to the manufacturer's instructions. RNA was labeled (biotin), fragmented, and hybridized to the arrays according to the manufacturer's recommendations.