NC和Lipofectamine®2000转染试剂,室温静置5 min,将NC与miR-140-5p mimics分别与转染试剂混匀后静置20 min;静置期间,将6孔板里的旧培养基弃掉,用PBS清洗细胞2次,加入800 μL新鲜DMEM培养基(不含血清和双抗);静置结束后将500 μL转染混合液均匀滴加到培养板中,轻轻晃动培养板使溶液混合均匀;培养箱中静置6 h...
进一步的双荧光素酶报告基因实验结果显示,野生型HDAC报告质粒(wt-HDAC4-luc)和 miR-140-5p mimics 共转染后,MG-63细胞的荧光素酶活性比wt-HDAC4-luc 和NC mimic组共转染组明显降低 (P<0.05);但将理论上的HDAC4 3′UTR与 miR-140-5p 的结合位点做了突变后(共转染wt-HDAC4-luc与miR-140-5p mimics),荧...
miR-140-5p mimics could downregulate the wnt1 mRNA and protein levels in MCF-7 and MDA-MB-231 cells. Furthermore, miR-140 mimics could enhance the sensitivity of BCSCs to doxorubicin (Dox) through the Wnt1/ABCB1 pathway both in vitro and vivo. Our findings have presented a novel miRNA...
The next day, the cells were transfected with 50 nM of miR-140-5p mimics or negative control mimics (Thermo Fischer) using Dharmafect1 (Thermo Scientific) following the manufacturer’s protocol. Simultaneously, the cells were transfected with luciferase constructs at a concentration of 100 ng per...
8、本发明首先通过对e14天孕鼠大脑皮层的神经干细胞进行体外培养发现mir-141-5p在神经干细胞成熟过程中呈现显著降低的趋势;之后采用mir-141-5p mimics验证了在神经干细胞中过表达mir-141-5p会显著减少神经干细胞的增殖能力,这说明抑制mir-141-5p表达的物质可以促进神经干细胞的增殖。
2.5 Transwell共培养体系中miR-155过表达/低表达对巨噬细胞IL-6、IL-10表达水平的影响Western blot法检测结果显示,miR-155 mimics组IL-6表达水平随时间进展呈上升趋势,IL-10表达水平呈下降趋势,在24 h时间点,miR-155 mimics组IL-6...
146a-5p agomir组,孕第16.5天(day 16.5,D16.5)对小鼠宫内总胎数/胚胎数、吸收胚胎数、存活胎数及存活胎鼠重量和胎盘重量进行计量分析;④分离培养原代小鼠骨髓源性巨噬细胞(bone marrow-derived macrophages,BMDM),利用LPS刺激诱导其M1...
miR-146a-5p 对猪骨骼肌卫星细胞(MuSCs)的影响:研究人员成功提取猪 MuSCs 后发现,miR-146a-5p 能抑制 MuSCs 增殖,促进其分化。miR-146a-5p mimics 可降低细胞增殖相关蛋白和基因表达,提高分化相关基因和蛋白表达;而 miR-146a-5p 抑制剂则产生相反效果34。
(miR-143), hsa-miR-145-5p (miR-145) and miR-negative (miR-neg, non-targeting control) miRNA mimics were purchased from Ambion (Applied Biosystems, Thermo-Fisher Scientific). Lipofectamine RNAiMAX (Invitrogen, Thermo-Fisher Scientific) was used for the transfection of the miRNA mimics according...
miR-708-5p mimics (mimics), miR-708-5p mimics control (nc), miR-708-5p inhibitor (inhibitor), and miR-708-5p inhibitor control (inhibitor nc) were purchased from GenePharma (Suzhou, China). The sequences were as follows: mimics Forward: 5′-AAGGAGCUUACAAUCUAGCUGGG-3...