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DDX39B interacts with the pattern recognition receptor pathway to inhibit NF-κB and sensitize to alkylating chemotherapy

机译:DDX39B与模式识别受体途径相互作用以抑制NF-κB并对烷基化化疗敏感

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摘要

DDX39B inhibits NF-κB activity. Luciferase assay using a reporter bearing -1C κB-sites in A172 cells expressing two independent sh-DDX39B constructs or a control vector. Data show mean value normalized to EV, ± SEM from two independent experiments. Luciferase assay in U87 cells transfected with empty vector (EV) or DDX39B. Data show mean value normalized to EV, ± SEM from two independent experiments. Immunoblot (IB) in A172 GBM cells expressing two sh-DDX39B constructs or sh-control. IB was performed with anti-phospho-IκBα, anti-IκBα, or anti-GAPDH as loading control. IB in GBM34 and GBM44 GSCs probed with anti-DDX39B. Immunoblot in GBM44 GSCs expressing shRNA constructs as in . IB performed as in . Representative immunofluorescence (IF) analysis of endogenous p65 in A172 cells expressing sh-DDX39B. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI). Scale bar, 10 μm. IB in GBM44 GSCs from expressing shRNA constructs probed with anti-phospho-p65 and anti-p65 antibodies. IB in GBM34 GSCs stably expressing empty vector (EV) or S-tagged DDX39B with anti-phospho-p65 and anti-p65. *  P t test)
机译:DDX39B抑制NF-κB活性。使用表达两个独立的sh-DDX39B构建体或对照载体的A172细胞中带有-1CκB位点的报告基因进行荧光素酶测定。数据显示两个独立实验的平均值标准化为EV,±SEM。空载体(EV)或DDX39B转染的U87细胞中的萤光素酶测定。数据显示两个独立实验的平均值标准化为EV,±SEM。表达两个sh-DDX39B构建体或sh-control的A172 GBM细胞中的免疫印迹(IB)。用抗磷酸-IκBα,抗-IκBα或抗GAPDH作为负载对照进行IB。用抗DDX39B探测的GBM34和GBM44 GSC中的IB。表达于shRNA的GBM44 GSCs中的免疫印迹。 IB的执行与表达sh-DDX39B的A172细胞中内源性p65的代表性免疫荧光(IF)分析。细胞核用4',6-二-2-基-2-苯基吲哚(DAPI)复染。比例尺,10μm。 GBM44 GSC中的IB来自表达的用抗磷酸化p65和抗p65抗体探测的shRNA构建体。 GBM34 GSC中的IB稳定表达带有抗磷酸化p65和抗p65的空载体(EV)或带有S标签的DDX39B。 * P t测试)

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