We have engineered a self-inactivating retroviral vector containing multiple (3 or 6) nuclear factor of activated T-cell (NFAT)-binding sites, followed by the minimal IL2 promoter and the reporter gene GFP. Jurkat cells, primary T-cell blasts, and T-cell clones were transduced with high ...
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建立IL2启动子以及NFATAP1增强子调控报告基因包括d2EGFP或Luciferase在Jurkat细胞中瞬时表达的细胞模型.首先,利用三步连接将IL2promoter-255～+285处的序列插入到pNFκBd2EGFP表达载体启动子上游,形成3个拷贝的前后串联的增强子序列;然后从人外周血钓取两条IL2promoter序列,包括-326～+46以及-89～+46两段基因组序列,分...
Finally, we tested the effect of the GFP-SPRIEIT fusion proteins on NFAT-activated gene expression in Jurkat T cells. Plasmids encoding GFP, GFP-SPRIEIT, and GFP-SPAIAIA proteins were introduced into Jurkat cells together with an NFAT-driven luciferase reporter plasmid, and a day later the ...
To assess the effects of NFATc1 activity on Notch- induced Ptcra promoter activation, 1 Â 106 Jurkat cells were transfected by the ViaFect Transfection Reagent (Promega, Wisconsin, USA) with 5 mg DNA containing all necessary constructs according to the manufacturer's protocol. Twenty four ...
NFAT-induced FasL expression has been implicated in Jurkat T, breast cancer, cardiac, neuronal, and Leydig cell apoptosis49–52, but whether NFAT regulates cell death induced by the FasL/Fas pathway in GMCs was unclear. Data here show that TRPC6 activation elevates membrane FasL level and ...
实验一:在上述不同时间点使用P/I进行刺激,MTT法检测Jurkat T淋巴细胞增殖,ELISA检测细胞上清液中细胞因子IL-2,IL-4,IFN-γ的表达,分析时间效应关系;同时采用流式细胞术Fluo-3AM检测胞浆钙离子水平,Western blot法检测CaN的改变,ELISA法检测NFAT活化情况,观察调节Mfn2的表达对Ca2+-CaN-NFAT信号通路各个环节的影响...
方法:体外培养人T细胞淋巴瘤细胞系Jurkat细胞,用含10%胎牛血清的 RPMll640培养基调整细胞浓度为1×106/ml,然后接种于6孔板,对其进行慢病 毒转染后进行效果验证,符合要求后进行分组,在Oh,6h,12h,24h,48h时间点处用 浓度为50ng/ml PMA/luM Ionomycin进行刺激。实验一:在上述不同时间点使用 P/I进行刺激,MTT法...
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初步筛选建立IL-2启动子以及NFAT-AP1增强子调控报告基因包括d2EGFP或Lueiferase在Jurkat细胞中瞬时表达的细胞模型.首先,利用三步连接将IL-2 promoter-255～+285处的序列插入到pNF-κB-d2EGFP表达载体启动子上游,形成3个拷贝的前后串联的增强子序列;然后从人外周血钓取两条IL-2 promoter序列,包括-326～+46以及-89...