small RNAnext Generation Sequencing of bone marrow-derived macrophages treated in M-CSF withdrawal medium
The goal of this study is to investigate the presence of novel small RNAs in apoptotic macrophage Overall design: Method: cultured mouse (bl6 line) bone marrow were differentiated in for 7 days in conditioned medium, in preence of Macrophage-Colony Stimulating Factor (M-CSF). M-CSF also stimulates the survive of macrophages by activating PI3K/AKT pathway. In accordance to previous publication (Lombardo, E., Alvarez-Barrientos, A., Maroto, B., Bosca, L., and Knaus, U.G, 2007, TLR4-mediated survival of macrophages is MyD88 dependent and requires TNF-alpha autocrine signalling, J Immunol 178, 3731-3739) apoptosis was induced in culturaed bone marrow-derived macrophages upon 36 hr of M-CSF withdrawal. Total RNA was isolated and a small RNA library was made using "Small RNA sequencing kit" (Illumina), according to the manufacturer's instructions.