Transcriptomics of PLXDC2 knockdown BMDM treated with PTEN. Transcriptomics of PLXDC2 knockdown BMDM treated with PTEN

To evaluate whether PLXDC2 mediated the role of PTEN on macrophages, PLXDC2 was knocked down in M2-polarized BMDMs, and the resultant PLXDC2-knocked down cells together with control cells were treated with or without PTEN, before subjected to global transcriptome profiling. Overall design: Bone-marrow (BM) cells were harvested from femurs and tibia of C57BL/6J mice. Differentiation was induced by recombinant mouse M-CSF in DMEM medium containing 1% penicillin/streptomycin and 10% FBS for 7 days to obtain the bone marrow-derived macrophages (BMDMs).To induce an M2-like phenotype, we treated BMDMs with IL-4 and IL-13 for 48 h.M2 BMDMs transfected with siNC or siPLXDC2#1,2 were treated with and without 100 ng/mL PTEN for 24 h, followed by RNA-seq analysis.

为探究PLXDC2是否介导PTEN在巨噬细胞中的调控作用，研究人员在M2极化的骨髓源性巨噬细胞（bone marrow-derived macrophages, BMDMs）中敲低PLXDC2，随后将得到的PLXDC2敲低细胞与对照细胞分别经PTEN处理或不做处理，再开展全转录组谱分析。实验整体设计：从C57BL/6J小鼠的股骨与胫骨中采集骨髓（bone-marrow, BM）细胞；将采集的骨髓细胞置于添加1%青霉素/链霉素与10%胎牛血清（fetal bovine serum, FBS）的DMEM培养基中，加入重组小鼠巨噬细胞集落刺激因子（M-CSF）诱导分化7天，以获得骨髓源性巨噬细胞（BMDMs）。为诱导M2样表型，使用IL-4与IL-13处理BMDMs 48小时。将转染阴性对照小干扰RNA（siNC）或靶向PLXDC2的小干扰RNA#1、#2的M2型BMDMs分别用100 ng/mL PTEN处理或不处理，持续24小时后进行RNA测序（RNA-seq）分析。