为了深入研究 MECP2 作为转录因子的功能,首要任务是明确其与 DNA 的结合位点。当前,通过传统的 ChIP-Seq 技术在基因组中精确定位 MECP2 与 DNA 的结合位置存在挑战,这主要是由于技术的分辨率限制,难以精确解析MECP2在DNA中的识别位点...
有意思的是,MeCP2可以作为脚手架蛋白负责这一复合物的组装,数据还提示了这一复合物的形成受到DNA甲基化与新生RNA转录的调控;在MeCP2 KO和T158M小鼠中,MeCP2/Rbfox/LASR复合物发生解离。接着,作者利用RNA-seq、iCLIP-seq、in v...
Here, we performed chromatin immunoprecipitation followed by sequencing (ChIP-seq) of Flag-tagged-H1.0 in mouse forebrain excitatory neurons. Unexpectedly, Flag-H1.0 and MeCP2 occupied similar genomic regions and the Flag-H1.0 binding was not changed upon MeCP2 depletion. Furthermore, mild ...
Our results of miRNA chip assay and ChIP-seq showed that MeCP2 inhibited the expressions of numerous miRNAs by binding to their upstream elements, including not only the promoter but also the distal enhancer. Among the affected miRNAs, miR-22 was identified to remarkably suppress gastric cancer ...
To assess the MeCP2 binding density, ChIP-seq reads were mapped to human (hg19) or mouse (mm10) genomes using Bowtie2 v2.2.3 default settings. MAC2 (ref.59) was applied to call MeCP2 peaks from mapped ChIP-seq reads, with an extension size of 250nt and aq-value (FDR) cut-off...
Through microarray analysis and chromatin immunoprecipitation sequencing (ChIP-Seq), we found that MeCP2 promoted GC cell proliferation by inhibiting FOXF1-mediated Wnt5a/β-Catenin signaling pathway and inhibited GC cell apoptosis by suppressing MYOD1-mediated Caspase-3 signaling pathway. 2. Materials...
Single-cell RNA-seq libraries were prepared by Chromium controller (10x Genomics) using Single Cell 3′ Reagent Kit v2 according to the manufacturer’s instructions. For each group, 17,400 cells were applied to a single lane of the Single Cell A Chip targeting 10,000 cells recovery. Paired-...
The percent methylated or hydroxymethylated DNA/ input DNA was calculated as described for ChIP. Amplicon-specific TAB pyrosequencing The distribution of 5-hmC and 5-mC in the GAD1 promoter amplicon was determined by a modification of TAB-seq.30 We followed the method originally described in ...
To determine the genome-wide distribution of MeCP2, nucleosomes and H1, we performed H1 chromatin immunoprecipitation sequencing (ChIP-seq) using a pan H1 antibody (Supplementary Fig. 4A) and compared the results to those of a high-resolution MeCP2 ChIP-seq and MNase-seq data sets13 (Fig....