PRM of DMXAA-treated Raw264.7 secretome

Raw264.7 cells were cultured in serum-free DMEM containing DMSO or DMXAA for 4 h, and the conditioned media were collected. The precipitated proteins were directly digested with trypsin/Lys-C mix. Three biological replicates of the samples were individually prepared and analyzed by targeted LC-MS/MS using parallel reaction monitoring (PRM).

Raw264.7细胞于含二甲基亚砜（Dimethyl sulfoxide，DMSO）或DMXAA的无血清杜氏改良伊格尔培养基（Dulbecco's Modified Eagle Medium，DMEM）中培养4小时，收集条件培养基。沉淀得到的蛋白质直接以胰蛋白酶/赖氨酸C混合酶进行酶解。对三份独立制备的生物学重复样品，采用靶向液相色谱-串联质谱（liquid chromatography-tandem mass spectrometry，LC-MS/MS）结合平行反应监测（parallel reaction monitoring，PRM）技术分别进行分析。