small RNAnext Generation Sequencing of bone marrow-derived macrophages treated in M-CSF withdrawal medium. Mus musculus

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.

本研究旨在探究凋亡巨噬细胞中新型小RNA的存在情况。
实验设计：
方法：将bl6品系小鼠的骨髓细胞在含有巨噬细胞集落刺激因子（Macrophage-Colony Stimulating Factor, M-CSF）的条件培养基中分化培养7天。巨噬细胞集落刺激因子还可通过激活PI3K/AKT通路促进巨噬细胞存活。参照此前发表的文献（Lombardo E、Alvarez-Barrientos A、Maroto B、Bosca L、Knaus UG，2007年，《Journal of Immunology》178卷，3731-3739页：TLR4介导的巨噬细胞存活依赖MyD88且需要TNF-α自分泌信号通路），在撤除巨噬细胞集落刺激因子36小时后，于培养的骨髓来源巨噬细胞中诱导细胞凋亡。依照制造商的操作说明，使用Illumina「小RNA测序试剂盒」完成总RNA提取与小RNA文库构建。