3. Barnes, Claire, and Aditi Kanhere. "Identification of RNA–protein interactions through in vitro rna pull-down assays." Polycomb Group Proteins: Methods and Protocols (2016): 99-113. 4. Hellman, Lance M., and Michael G. Fried. "Electrophoretic mobility shift assay (EMSA) for detecting p...
ALKBH5的功能之一是RNA去甲基化,那么它是否是通过去甲基化影响癌细胞的表型呢?作者首先通过dot blot assays检测了细胞中m6A的总体水平(图4A),同预期一致,ALKBH5缺失使得细胞中m6A修饰的总体水平上升,而ALKBH5过表达细胞中m6A修饰总体水平明显下降。 图4 为了找出依赖...
5、miR-145通过调节YTHDF2调控mRNA的水平:在HepG2细胞中转染pcDNA3.1-YTHDF2,通过dot blot assays、RT-PCR和IHC技术,发现过表达YTHDF2后mRNA的水平降低。在HepG2细胞中单独转染miR-145,mRNA的水平显著上升;共转染miR-145和pcDNA3....
(1)使用商业化的细胞增值检测盒(CCK-8、EdU)来评估PC细胞的增殖活性(图2/a,e),结果显示ALKBH5的过表达可以显著地减少PC细胞的增殖;利用菌落形成试验(Colony formation assays)也可以观察到ALKBH5的过表达削弱了PC细胞菌落的长期形成,ALKBH5的敲除则呈现了相反的结果; (2)作者将流式细胞术和WB实验进行结合(图2...
Transcriptome-wide m6A-seq and RNA-seq assays identified potential targets of METTL3 in colorectal cancer. (a) The m6A dot blot assay of global m6A abundance in mRNA of HCT116 WT and METTL3-knockout cells. MB, methylene blue staining (as a loading control). (b) The global m6A levels ...
The global m6A modification level was detected by dot blot assays. Our results confirmed that m6A modification was indeed upregulated in OXA-R tissues, NASH-HCC tissues and OXA-R cells (Fig. 5A). To evaluate m6A modification level in steatosis, an oleate-treated HepG2 cell model was ...
a m6A dot blot assays showing global m6A level of RNAs extracted from 40 pairs of GC tissues. RNAs were serially diluted and loaded equally with the concentration of 50 ng/μl. The methylene blue staining (below) was used to detect input RNA, while the intensity of dot immunoblotting (...
Early studies explored m6A modification in RNA by radiolabeling methods such as radioisotope labeling [7]. In addition, enzyme-linked immunosorbent assays (ELISA) utilize anti-m6A antibodies to detect m6A levels in RNA, mainly for quantitative analysis. Dot blot is a rapid method for detecting m6A...
m6A dot blot assays m6A dot blotting was performed as previously described [20]. Total RNA was harvested as previously described with RNA quantity monitoring. RNA was denatured by heating at 95 °C for 10 min and then preserved on ice. Then, mRNAs were blotted onto an N+ nylon membrane (...
(1)使用商业化的细胞增值检测盒(CCK-8、EdU)来评估PC细胞的增殖活性(图2/a,e),结果显示ALKBH5的过表达可以显著地减少PC细胞的增殖;利用菌落形成试验(Colony formation assays)也可以观察到ALKBH5的过表达削弱了PC细胞菌落的长期形成,ALKBH5的敲除则呈现了相反...