本研究中,作者们以STAT3双位点磷酸化在胃癌细胞发生发展的重要角色为背景,基于STAT3双位点磷酸化在肿瘤细胞中介导不同生物学功能建立了全新的STAT3双磷酸化抑制剂的高通量筛选实验,即表征STAT3 p-Tyr705抑制活性的STAT3依赖性荧光素酶实验(Luciferase reporter assay)和用于表征p-Ser727抑制活性的ATP抑制实验 (ATP inhib...
本研究中,作者们以STAT3双位点磷酸化在胃癌细胞发生发展的重要角色为背景,基于STAT3双位点磷酸化在肿瘤细胞中介导不同生物学功能建立了全新的STAT3双磷酸化抑制剂的高通量筛选实验,即表征STAT3 p-Tyr705抑制活性的STAT3依赖性荧光素酶实验(L...
Successful silencing and inhibition of phosphorylation was confirmed using Western blot analysis and a luciferase reporter assay. RNAi‐mediated silencing as well as STATTIC treatment resulted in significantly decreased clonogenic survival following exposure to 3 M of 5‐FU and irradiation in a dose‐...
Dual-luciferase reporter assay The 293T cells were co-transfected with the IFN-β-Luc reporter, the pRL-TK and sh-STAT3 plasmids, or control sh-luciferase plasmid for 24 h, and then infected with SeV for 16 h. Luciferase assays were performed using the dual-luciferase activity assay kit ac...
Luciferase reporter assay The Fam3a regulatory region (−3000 to +100 bp) was cloned into a pGL3 Luciferase plasmid (Fam3a-Luc). The following vectors were used: Fam3a-Luc, pRL Renilla Luciferase Control Reporter Vector (Promega), pcDNA3-Stat366, and pcDNA3-myc-MyoD41. HEK293 cells ...
Luciferase reporter assay was used to confirm the direct targeting of microRNAs. Gelatin zymography was used to study matrix metalloproteinase (MMP) activity. Transwell assay was used to investigate cell migration and invasion. Results: Enforced expression of STAT3 decreases the endogenous MMP inhibitor ...
[72]. In apoptotic MC3T3-E1 cells under iron overload conditions, the expression of miR-3074-5p was upregulated. The results of the dual-luciferase reporter assay showed that overexpression of miR-3074-5p caused apoptosis by regulating its downstream target gene Smad4, which blocked the ...
Luciferase Reporter Assay For the reporter assays, 1000-2000 cells were seeded in 384-well white plate (Corning Costar). Two duplicate plates were set up: one for luciferase reporter assay and one for CellTiter-Glo cell viability assay. Four to six hours later, drugs were added to each well...
将细胞系以1×104个细胞的密度接种于96孔板中,第二天用fugenehd(promega)转染25ngtk-renilla质粒作为内参。细胞转染24h后加入不同浓度的imp预处理4h,再加入tcm刺激24h,使用dual-luciferasereporterassaysystem(promega)检测荧光值,结果如图8c所示。imp在5-20μm浓度下能够剂量依赖的抑制stat3的转录活性。
Then, the cells were collected and lysed in passive lysis buffer (Promega) for 15 min, the transcriptional activity of STAT3 was determined using a luciferase reporter assay system (Promega), according to the manufacturer’s instructions. EMSA A number of 4 × 105 U266 cells were ...